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Enterobacter cloacae complex isolates for antibiotic resistance studies

Enterobacter cloacae is part of the normal human flora and not considered a primary pathogen. Unfortunately, there is increasing incidence of nosocomial infections with E. cloacae, which frequently involve intravenous and other medical and hospital devices as well as medical solutions and disinfectants used routinely in hospitals and patient care facilities. The situation is further complicated by the ability of these bacteria to acquire resistance to antibiotics, notably carbapenems. BEI Resources has obtained a set of 15 isolates of the E. cloacae complex with defined susceptibility patterns against 16 antibiotics. This valuable set is now available to aid your studies on carbapenem-resistant Enterobacteriaceae (CRE).

BEI Resources No.	Description
NR-50391	Enterobacter cloacaecomplex strain BEI01
NR-50392	Enterobacter cloacaecomplex strain BEI02
NR-50393	Enterobacter cloacaecomplex strain BEI03
NR-50394	Enterobacter cloacaecomplex strain BEI04
NR-50395	Enterobacter cloacaecomplex strain BEI05
NR-50396	Enterobacter cloacaecomplex strain BEI06
NR-50397	Enterobacter cloacaecomplex strain BEI07
NR-50398	Enterobacter cloacaecomplex strain BEI08
NR-50399	Enterobacter cloacaecomplex strain BEI09
NR-50400	Enterobacter cloacaecomplex strain BEI10
NR-50401	Enterobacter cloacaecomplex strain BEI11
NR-50402	Enterobacter cloacaecomplex strain BEI12
NR-50403	Enterobacter cloacaecomplex strain BEI13
NR-50404	Enterobacter cloacaecomplex strain BEI14
NR-50405	Enterobacter cloacaecomplex strain BEI15

Mayaro and Una Viruses Now Available

Mayaro virus (MAYV) is a New World alphavirus and the etiologic agent of Mayaro fever, an acute febrile illness sometimes accompanied by severe and persistent arthritis similar to Chikungunya fever. Its transmission cycle occurs mainly through mosquito vectors, especially those of the genus Haemagogus, but Aedes spp. mosquitoes may also be competent vectors. The enzootic transmission cycle of MAYV is not fully understood, but the occurrence of relatively large outbreaks of Mayaro fever and the competence of Aedes mosquitoes for the transmission of MAYV suggest the potential for an urban human-mosquito-human transmission cycle to emerge. MAYV was first isolated in Trinidad in 1954, with sporadic outbreaks of Mayaro fever in Bolivia, Brazil, and other regions throughout Central and South America. Recently, MAYV was found in a DENV co-infected patient in Haiti¹, the first report of MAYV in the Caribbean nation, underscoring the need for MAYV inclusion in surveillance activities in these regions in addition to Dengue, Chikungunya, and Zika viruses.

Una virus (UNAV) is the closest genetic relative of MAYV that also causes a similar febrile illness. UNAV was first identified in northern Brazil in 1959. Serosurveys have identified antibodies against UNAV in birds, horses, and humans; however, the extent of viral distribution and human disease risk is unknown.

References:1. eid2211.161015

Plasmid stability in Staphylococcus without antibiotic selection?

Plasmid maintenance by bacteria can be unpredictable without selective pressure. This is of particular concern during in vivo studies. To circumvent this in Staphylococcus species, the Bose Lab at The University of Kansas Medical Center has provided plasmids to BEI Resources that remain stable inStaphylococcus cells without the requirement of antibiotics during both in vitro and in vivo experiments¹. These E. coli-S. aureus shuttle vectors are based on the LAC-p01 plasmid.

Plasmid pKK22 (Accession KX085042) is designed to be used with S. aureus USA300 strains that contain LAC-p01 and will render the strains isogenic. Plasmid pKK30 (Accession KX085043) is a variant of pKK22, in which the predicted open reading frames (ORFs) that are not needed for plasmid maintenance have been deleted, and is intended for use inStaphylococcus cells that do not contain LAC-p01. Both plasmids contain a single trimethoprim resistance cassette that is functional in both E. coliand Staphylococcus. In addition, they contain the E. coli R6Kγ origin of replication and require pir+ cells for replication. DH5αλpir and the pir+dam- dcm- strain GM2163λpir are provided as a host strains, courtesy of Dr. Eric Stabb (University of Georgia)². These plasmids and host strains are now supplied individually or as a kit through BEI Resources.

BEI Resources No.	Description
NR-50348	Escherichia coliDH5αλpir containing pKK22
NR-50349	Escherichia coliDH5αλpir containing pKK30
NR-50350	Escherichia coli DH5αλpir
NR-50351	Escherichia coli GM2163λpir
NR-50352	Kit: In vivo stable Staphylococcus plasmids and E. coli hosts

References:

1. Krute, CN, KK Krausz, MA Markiewicz, JA Joyner, S Pokhrel, PR Hall, and JL Bose. 2016. Generation of a stable plasmid for in vitro and in vivostudies of Staphylococcus. Applied and Environmental Microbiology.82:6859-6869. PMID 27637878

2. Dunn AK, MO Martin, and EV Stabb. 2005. Characterization of pES213, a small mobilizable plasmid from Vibrio fischeri. Plasmid54:114-134. PMID: 16122560

Clostridium difficile research is easier with strains from BEI Resources

Clostridium difficile research is easier with strains from BEI Resources

Clostridium difficile (also known as Peptoclostridium difficile) is an increasingly common opportunistic pathogen that causes diarrhea and more serious intestinal conditions such as colitis. It is commonly observed in patients that have an illness that requires prolonged use of antibiotics, or can be acquired in hospitals and long-term care facilities. C. difficile infections are normally treated with antibiotics; however, this is particularly problematic due to the high levels of antibiotic resistance displayed by these bacteria. Recent studies have shown that replenishment of the normal gut microbiota provides a successful tool to control the infection with C. difficile.
BEI Resources has a large number of C. difficile strains, as well as nucleic acids, toxins, and other reagents available for your research (please click here for a list).
Among the bacterial strains, several have well characterized tcdA, tcdB, and cdtB toxin

Viral nucleic acids

For the benefit of researchers requiring nucleic acids from pathogens, BEI Resources offer more than 1,000 nucleic acids derived from pathogens. These reagents can be used at lower containment levels for molecular studies; to ensure their biosafety, the preparations have been safety tested for the absence of residual live organisms. Please check our website to see the wide variety viral nucleic acids extracted from various BSL-2 through BSL-4 pathogens. Notable new additions to this category of reagents include Genomic RNA from Enterovirus D68 and human metapneumovirus.

Give a whooping push to your research

Whooping cough is a disease caused by bacteria from the genus Bordetella. Currently under widespread resurgence, whooping cough ranks globally among the 10 leading causes of childhood mortality. BEI Resources offers several isolates of Bordetella pertussis, B. bronchiseptica, and B. holmesii, along with toxins, nucleic acids, and proteins for Bordetella research. To see the full list of Bordetella reagents and strains currently available from BEI resources, please click here

Image: Bordetella holmesii NR-43499 grown on Bordet Gengou media

Toxoplasma gondii

Toxoplasma gondii is an obligate intracellular protozoan parasite of the phylum Apicomplexa that is the causal agent of toxoplasmosis. BEI Resources houses a comprehensive assortment of T. gondii cultures that include GFP-expressing strains; reference genome strains; genetic crosses between type strains I, II, and III; and strains used as models for parasite transmission and cyst formation. For a complete list of our Toxoplasma catalog, please contact us at Contact@BEIResources.org.

BEI Resources Number	Comment	Strain
NR-743	Transgenic RH strain expressing GFP	RH-GFP 5 S65T
NR-744	Transgenic PTG strain expressing GFP	PTG-GFP 5 S65T
NR-10150 - NR-10170	Genetic crosses between strain types II and III	Various
NR-10238 - NR-10273	Genetic crosses between strain types I and III	Various
NR-49173 - NR-49199¹	Genetic crosses between strain types I and II	Various
NR-20728 - NR-20739	Reference genome strains for haplogroups 1 through 15	Various
NR-15248	Mutant RH strain deficient in the ku80 and hxgprt genes	RH∆ku80∆hxg
NR-43998	Pig, Massachusetts, 2002; cat-to-cat transmission studies	Dubey
NR-44106	Recombinant type I/III; forms cysts spontaneously in vitro	EGS

¹ Available soon. Send inquiries to Contact@BEIResources.org for availability.

Staphylococcus gene expression analysis shuttle vectors for pathogenesis studies

BEI Resources offers 42 distinct Escherichia coli-Staphylococcus cassette-based shuttle vectors that provide all the tools you need to complete your research. These shuttle vectors are deposited as part of the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA) reagents. Choose from staphylococcal low-copy-number, high-copynumber, thermosensitive, and theta-replicating plasmids. Select from erythromycin, tetracycline, chloramphenicol, kanamycin, or spectinomycin selectable markers in Staphylococcus, and ampicillin, erythromycin, kanamycin, and chloramphenicol resistance markers in E. coli. The modular design of the plasmid vector system offers flexibility and variety for expression analysis and provides the opportunity to address questions about gene dosage, complementation, and cis-trans effects. The shuttle vector system provides effective tools for studying gene regulation of staphylococci among ecosystems.

For further published details on these plasmids, please see Charpentier et al. Appl

Environ Microbiol 70(10): 6076-6085, 2004

Image: NR-46158. Staphylococcus aureus strain 4221 containing shuttle vector pCN57, grown on TSA + 10 μg/ml erythromycin

Genome Sequenced Clinical and Contaminated Yogurt Isolates of Mucor circinelloides f. circinelloides

Two genome sequenced isolates of Mucor circinelloides f. circinelloides (NR-49108 Strain designation 1006PhL, and NR-49117 Strain designation Mucho) are now available from BEI Resources. Mucor circinelloides f. circinelloides is surpassed only by Rhizopus species as the most common cause of mucormycosis, and is the most virulent of the Mucor circinelloides species. One isolate was obtained from the skin of a healthy individual, and the other from contaminated yogurt responsible for the 2013 Mucorales outbreak.

Filoviral expression plasmids

We have a number of expression plasmids for various Filoviral proteins for Zaire Ebolavirus and Marburgvirus. Many of the proteins have been tagged with HA or FLAG. Continue to check our catalog for new plasmid additions, such as those expressing GP, NP, VP24, VP30, VP35, and VP40 in the next few months for a complete list.

Pathogenic amoebae that affect human health

Staphylococcus aureus

Entamoeba histolytica

Pathogenic amoebae that affect human health

Amoebae are a diverse group of protists that move primarily by extending and retracting pseudopodia. The most important pathogenic amoeba for humans is Entamoeba histolytica. Among the free-living amoebae that exist in nature, Acanthamoeba spp. and Naegleria fowleri belong to a handful of genera that have an association with human disease. BEI Resources houses a diversity of pathogenic amoeba strains for researchers working on mechanisms of pathogenesis, diagnostics, antimicrobial sensitivity, and the molecular biology of these organisms.

Entamoeba histolytica. Entamoeba histolytica is an enteric parasite that predominantly infects humans and other primates. Most infections are asymptomatic and tissue invasion is a rare occurrence. In some cases, diseases can range from chronic, mild diarrhea, to fulminant dysentery. For a list of E. histolytica strains available at BEI Resources click here.

Acanthamoeba spp. Acanthamoebae inhabit a wide variety of environmental niches. In healthy humans, Acanthamoeba can cause a sight-threatening eye disease known as Acanthamoeba keratitis. In immunocompromised individuals, Acanthamoeba can cause disseminated infections and the fatal disease granulomatous amebic encephalitis. For a list of Acanthamoeba spp. strains available at BEI Resources click here.

Naegleria fowleri. Naegleria fowleri is the causative agent of primary amoebic meningoencephalitis (PAM) in humans. The disease is rare yet so fulminant that few people survive infection. Cases of PAM reported in North America usually occur after swimming in water naturally heated by the sun. For a list of Naegleria fowleri strains available at BEI Resources click here.

New Updates to MR4 Methods Manuals

Anopheles mosquito. Photo courtesy of James Gathany and CDC

Anopheles mosquito. Photo courtesy of James Gathany and CDC.

New Updates to MR4’s “Methods in Anopheles Research” and “Methods in Malaria Research” Laboratory Manuals - Now Available for Free Download

Our friends at the CDC have updated the frequently-accessed and freely available MR4 Methods in Anopheles Research laboratory manual, which provides detailed insectary and benchtop protocols for rearing, genotyping, and phenotyping Anopheline and other mosquito vector colonies, as well as information on field techniques, insecticide resistance, screening for Wolbachia infection, and an introduction to Aedes and Culex colony methodology. To access the updates, download the full manual at no cost.

What’s more, a revised and expanded edition of EVIMalR and MR4’s extremely popular “Methods in Malaria Research” laboratory manual was also recently released. The updated manual provides a wide variety of protocols for in vitro, in vivo, antimalarial drug screening, and transmission research on Plasmodium spp., and the use of parasite genome databases to incorporate bioinformatics. Download a free, PDF version of this manual to access the latest updates.

Transposon library mutants available

Transposon Library Mutants

NR-46829. Staphylococcus aureus JE2 transposon mutant SAUSA300_0113 grown on TSB + 5 µg/mL erythromycin

Transposon library mutants available to study non-essential genes of Staphylococcus aureus

As part of the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA), BEI Resources is supplying the scientific community with the Nebraska Transposon Mutant Library (NTML). The NTML is an assortment of sequence-defined transposon insertion mutants of S. aureus strain USA300 LAC. Each clone harbors a different non-essential gene disrupted by the insertion of the mariner-based transposon bursa aurealis. The exact location of each transposon insertion in each of the 1955 clones is available from the University of Nebraska Medical Center. Most of these clones are grown when ordered (made to order), please allow time for production and testing by BEI Resources when ordering.

Human Metapneumovirus

Human Metapneumovirus

BEI Resources announces the addition of human metapneumovirus (hMPV) isolates, dating from 1983, to our vast assortment of viruses. hMPV was isolated from young children with acute respiratory tract disease in the Netherlands in 2001, and is now recognized as a major cause of respiratory illness in infants and children worldwide. hMPV is difficult to grow in cell culture, largely explaining the delay in recognizing this pathogen, which has been causing disease for half a century as evidenced by retrospective serological analyses. Please check our Knowledge Base query box using keyword “metapneumovirus” for tips on how to successfully propagate the virus in cell culture.

Welcome NARSA Researchers!

The NARSA Strain Repository is transitioning to BEI Resources, NIAID's centralized research reagent repository in May 2014. Due to this transition, Eurofins will no longer be registering users after March 14th, 2014. Eurofins will no longer be taking orders in May. All orders after May 23^rd, 2014, should be made through BEI Resources.

Registration for the BEI Resources program can be found at this link below. NARSA Strain requestors will need to register for Level 2 for continued access to the NARSA Strains. ** Please note if you are an existing NARSA registrant, BEI Resources will contact you after the Annual Meeting with specific registration instructions and paperwork to facilitate your registration.

http://www.beiresources.org/Register.aspx

Once registered with BEI Resources, researchers will have access to the full BEI Resources catalog (within the respective Biosafety Level). The catalog has over 12,000 research reagents representing more than 400 Genus and Species focusing on NIAID's Category A, B and C priority pathogen list and Emerging Infections.

Once registered with BEI Resources, ALL investigators will receive FREE shipping and handling on orders placed through BEI Resources.

Please feel free to contact BEI Resources by email: contact@beiresources.org or by phone: Toll-free telephone number: (800) 359-7370 (8:30 AM to 4:30 PM Eastern Time) with any questions or concerns.

Schistosoma spp. Life Cycle Training Course

General Information about the Schistosoma spp. Life Cycle Training Course

Biomedical Research Institute

12111 Parklawn Drive Rockville, MD.

May 21-23, 2014

This course is designed to acquaint investigators working in the field of schistosomiasis with basic techniques and strategies for maintaining Schistosoma mansoni, S. haematobium, and S. japonicum in the laboratory, and it offers directions into making one’s research more productive. Among the topics covered will be procedures for setting up and maintaining the complete S. mansoni/Biomphalaria glabrata life cycle in the research laboratory, an overview of schistosomiasis, the pathology of the disease, collection of adult worms and eggs from tissues, exposure of snails to miracidia, shedding snails, exposure of mice and hamsters to cercariae, and an examination of all the intramolluscan life stages. Hands-on exercises relating to some of these topics will also be presented (ex. perfusion of mice, exposing mice/hamsters, identification of sporocysts in snails). Since we first began offering this basic course, over 120 scientists, technicians and students have benefited from completion of the course, and have expanded their own research agendas once back in their own laboratories.

The workshop incorporates aspects of all three Schistosoma spp. life cycles. Attendees will be exposed to aspects of maintaining S. japonicum and S. haematobium life cycles that include general snails maintenance, collection of cercariae from infected snails, and exposing mice/hamsters. It is our hope that incorporating the other two life cycles will encourage interest in maintaining these species in attendees’ laboratories. The three-day course will also include a lecture on NIH-NIAID parasitology and grant programs.

We anticipate that attendees will come away with a greater appreciation of the basic biology of Schistosoma spp. parasites, greater confidence in maintaining life cycles, and they will learn strategies for optimizing life cycles in their home laboratories. Time will be available to ask questions about life cycle maintenance and other aspects of schistosomiasis research. There is no fee for the course. Attendees are responsible for travel and lodging expenses, but travel assistance may be available. If you have any questions about the course, please contact me at mtucker@afbr-bri.com or (301) 881-3300 x31, or Mitzi Sereno at msereno@afbr-bri.com or (301) 881-3300 x39.

MERS-CoV Now Available to Qualifed Labs

BEI Resources has the MERS-CoV virus (strain EMC/2012) available for distribution to qualified laboratories. This virus will require a BEI Level 3 registration. Middle East Respiratory Syndrome (MERS) is viral respiratory illness first reported in Saudi Arabia in 2012. It is caused by a coronavirus called MERS-CoV. Most people who have been confirmed to have MERS-CoV infection developed severe acute respiratory illness. They had fever, cough, and shortness of breath. About half of these people died.

The catalog number for the MERS-CoV virus is NR-44260. You must be logged in with a valid BEI Level 3 Account to place an order. Please email our customer service department if you have any questions about obtaining the MERS-CoV virus.

H7N9 Influenza Virus

The much awaited H7N9 Influenza virus is now available for your research needs. NR-44078 is a 6:2 reassortant virus produced using plasmid-based reverse genetics to contain the hemagglutinin and neuraminidase genes from A/Shanghai/1/2013 (H7N9) and internal genes from A/Puerto Rico/8/1934 (H1N1).

Click here to see all H7N9 reagents.

Upcoming Meetings & Events

Click Here to view our Upcoming Events Calendar.

New Website Redesign

As of Wednesday, August 20th we have updated the BEI Resources and MR4 websites to further optimize your online experience; making it easier, more accessible, and hopefully a bit more pleasurable to access your desired BEI and MR4 reagents. We have rolled out a new redesigned website that we anticipate will provide a more user-friendly navigation and shopping cart experience, easier searching and reading throughout the site, and an optimal viewing experience from essentially any web accessible platform whether it is desktop, laptop, mobile phone, or tablet.

This Responsive Web Design approach suggests that design and development should respond to the user’s behavior and environment based on screen size, platform, and orientation. As more and more of our customers in the scientific community are switching to the duality of laptops and mobile devices to conduct and support their research efforts, we at BEI Resources, would like to let our customers know… We’ve heard you… and would like to thank you for your continued feedback.

This redesigned website has been engineered to automatically respond to user preferences; switching to accommodate for resolution, image size and the scripting capabilities as dictated by the device being used. We can now make images and layouts more flexible. With this design, we have essentially eliminated the need to make individual adjustments in design and development for each new mobile device produced and made available on the market.

While Responsive Design is not a complete solution, we believe this solution provides more options for our users and a more optimal online experience overall when accessing the BEI Resources and MR4 sites; from any website accessible device; mobile or otherwise.

So feel free to access the BEI Resources and MR4 sites while in the office, lab, or on-the-go with your mobile device. An option to provide feedback is available through our Online User Feedback form, provided at the completion of your online order. Tell us what you think; we look forward to hearing from you.

At MR4 and BEI Resources, “We take care of the details - so you can focus on your research.”

SARS Virus Now Available for Researchers

We have available for the first time in the BEI Resources catalog, two SARS Coronaviruses (SARS-CoV) derived from the Urbani strain. NR-18925 is produced from a recombinant infectious clone, icSARS-CoV. NR-15418 is the mouse-adapted strain, Urbani v2163, developed by Dr. Barnard as a lethal model for evaluating antiviral agents (PubMed:19853271). To view a complete listing of all coronaviruses in the catalog, click here.

Sand Flies, Reduviids and Ticks are Here!

BEI Resources announces the addition of Sand Flies, Reduviids and Ticks for the research community.

Catalog Number	Vector	Pathogen Competence
NR-43999	Phlebotomus papatasi, PPNS, Larval	Leishmania major and Sand Fly Fever phleboviruses.
NR-44000	Phlebotomus papatasi, PPTK, Larval	Leishmania major and Sand Fly Fever phleboviruses.
NR-44001	Lutzomyia longipalpis, LLJB, Adult	Leishmania spp. including Leishmania infantum chagasi.
NR-44013	Phlebotomus papatasi, PPNS, Adult	Leishmania major and Sand Fly Fever phleboviruses.
NR-44014	Phlebotomus papatasi, PPTK, Adult	Leishmania major and Sand Fly Fever phleboviruses.
NR-44015	Lutzomyia longipalpis, LLJB, Adult	Leishmania spp. including Leishmania infantum chagasi.
NR-42510	Ixodes scapularis	Borrelia burgdorferi, Anaplasma phagocytophilum, Ehrlichia muris-like agent, Powassan virus, Babesia spp.
NR-42511	Ixodes ricinus	Anaplasma phagocytophilum, Tick-borne encephalitis virus, Borrelia burgdorferi, Babesia spp.
NR-42512	Rhipicephalus sanguineus	Rickettsia rickettsii, Rickettsia conorii, Ehrlichia canis
NR-42513	Dermacentor variabilis	Rickettsia rickettsii, Francisella tularensis, Colorado tick fever virus
NR-42514	Amblyomma americanum	Ehrlichia chaffeensis, Ehrlichia ewingii, Panola Mountain Ehrlichial agent
NR-44076 and NR-44077	Rhodnius prolixus	Trypanosoma cruzi

CDC recommends handling naïve ticks from these species at ABSL2 containment in appropriate barrier protected facilities; pathogen research may require elevated ABSL containment. Check the BEI Vector Resources page for a list of other vectors and related reagents available through BEI Resources.

Chikungunya Viruses - Now Available for Order

Chikungunya Viruses - Additional Strains Available

BEI Resources announces the addition of two novel BSL3 strains of Chikungunya virus (CHIKV), R-91142 and 182/25, to the catalog of available items. CHIKV is a mosquito-borne alphavirus that is mainly associated with acute febrile illness. Strains R-91142 and 182/25 CHIKV are of Asian origin. Strain 182/25 is the recently developed vaccine candidate and the only CHIKV strain to be tested in humans. Coming soon to the BEI catalogue is the CHIKV African prototype strain, S-27.

NR-13221	Chikungunya Virus, R-91142 (cell lysate)
NR-13222	Chikungunya Virus, 181/25 (cell lysate)
NR-13220	Chikungunya Virus, strain S-27 (cell lysate) *Coming soon!*

Resources for Trypanosome Research

Recent acquisitions in BEI Resources aim to contribute to research focused on understanding the mechanisms of disease and the development of novel therapeutic drugs for Trypanosome research. Such acquisitions include SSGCID clones, proteins and reference parasite strains used in genome and virulence studies. For more information on the availability of these materials please contact BEI Resources customer service.

Catalog Number	Organism	Strain	Comments
NR-36197	Trypanosoma brucei brucei	STIB 247	In vivo strain, causes chronic infection in mice
NR-36198	Trypanosoma brucei gambiense	STIB 386	In vivo strain, causes chronic infection in mice
NR-36630	Trypanosoma cruzi	TcVT-1	Isolated from chagasic dog in Virginia, USA
NR-40347	Trypanosoma cruzi	Y strain (+luc)	Luciferase-expressing transgenic strain
NR-41946	Trypanosoma brucei brucei	TREU 927/4 (GUTat 10.1)	Genome strain
NR-41947	Trypanosoma congolense	IL3000	Genome strain
NR-41948	Trypanosoma brucei rhodesiense	WRATat (stock LVH/75/USAMRU-K/18) (MVAT7)	Classic strain, isolated from human patient, Kenya, 1975
NR-42009	Trypanosoma brucei	Lister 427 VSG 221	Wild type bloodstream form
NR-42010	Trypanosoma brucei	Lister 427	Wild type procyclic form
NR-42011	Trypanosoma brucei	Lister 427 VSG 221 (TetR T7RNAP)	Transgenic bloodstream form co-expressing TetR and T7RNAP
NR-42012	Trypanosoma brucei	Lister 427 29-13 (TetR T7RNAP)	Transgenic procyclic form co-expressing TetR and T7RNAP

BEI Resources Fungi Repository is Growing!

BEI Resources is working to expand the fungal resources so that researchers have a broader range of organisms and reagents for their work. BEI Resources currently has eight genus and species represented with several more currently going through the production pipeline. BEI Resources is actively encouraging depositors with medically relevant strains to deposit into the repository. If you are interested in depositing, please click here.

Cloned lines of Toxoplasma gondii are now available from BEI Resources

Toxoplasma gondii is an obligate intracellular protozoan parasite of the phylum Apicomplexa that is the causal agent of toxoplasmosis. T. gondii is dominated by three widespread clonal lineages, referred to as types I, II, and III. The three major Toxoplasma lineages differ in a number of phenotypes, the best described of which is virulence in mice.

Recent studies have examined the genetic basis for these differences by mapping virulence in F1 progeny derived from crosses between the different T. gondii lineages.

BEI Resources houses various cloned lines selected from progeny of two parallel genetic crosses between a Type II parental strain (ME49 clone B7; NR-10150) and a Type III parental strain (CEP; NR-10151). Authentication of individual clones is performed through phenotypic and genotypic testing. This includes drug susceptibility to adenine arabinoside (ara-A) and sinefungin (Fig. 1) and PCR-based analysis of the SAG1 and KT-850 loci (Fig. 2).

Additional cloned lines from a Type I and Type III genetic cross will be available soon. In addition, BEI Resources offers more than 50 types of polyclonal antibodies to various Toxoplasma proteins. For technical questions about these products, or to learn more, please contact us by clicking here.

ATCC Adds ISO Guide 34:2000 and ISO 17025:2005 Accreditation to the ISO 9001:2000 Certification

ATCC (American Type Culture Collection), the contract manager of BEI Resources, has announced its accreditation for ISO Guide 34:2000 and ISO 17025:2005, an international multi-industry standard specifically designed for producers of reference materials.

The ISO Guide 34 accreditation by the American Association for Laboratory Accreditation (A2LA) builds upon the earlier certification for compliance with the ISO 9001:2000 standard for quality management systems received by ATCC in 2007. Biological reference materials produced under an ISO Guide 34-accredited process have confirmed identity, well-defined characteristics and an established chain of custody — qualities essential to their effectiveness as biological standards in research and development.

In gaining ISO recognition, ATCC joins a pioneering group of biological product organizations with ISO certification. Biological materials authenticated and preserved by ATCC, such as cell lines and microorganisms, are widely used as reference materials for research and product testing.

ATCC Senior Director for Quality, Compliance and Biosafety Barry Waters, PhD, remarked,“ISO 34 accreditation represents an objective measure of confidence in the consistency and quality of ATCC reference cultures. It expands ATCC’s ability to produce internationally recognized standards for biological research and development, including certified reference materials (CRMs).”

ISO 17025:2005 sets general requirements for the competence of laboratory testing and assures customers that the characterization and purity protocols used in the manufacture of products are precise, accurate and have repeatability. The scope of the accreditation includes specific tests or properties measured for microbial and cell cultures.

ISO 9001:2000 for quality management systems, ISO Guide 34 for reference material production and ISO 17025 for laboratory testing are among the standards promulgated by the International Organization for Standardization (ISO), a network of national standards institutes from 157 countries. ISO operates a Central Secretariat located in Geneva,Switzerland to coordinate the system.

The BEI Resources Website got a Make Over!

BEI Resources is pleased to announce a new and improved website! We heard your feedback concerning difficult searches and long checkout times…and, we invite you to explore and discover our new dynamic search package, state-of-the-art shopping cart experience and enhanced product information pages for your items on interest.

Our new search engine provides the community with a user friendly interface in which to research, search, and order reagents. We will now be using the same search and navigation features of many of the world's largest internet sites. Some of the major search features include dynamic filter searching, Intuitive Auto-Complete Text Searches, Suggested Search Terms, Dynamic Clustering and multiple sorting parameters for result sets.

Your ordering experience is also vastly improved with our new Web Services E-Commerce software. No more waiting for your items to load to the shopping cart; with the new software, your checkout is fast and reliable. Look for all these updates today!

Diarrheagenic E. coli organism panels

Outbreaks of foodborne disease caused by E. coli bacteria have become a serious problem in this country. E. coli organism panels, genomic DNA, and technical information are now available to support research to detect, treat, and prevent foodborne diseases.

In February of 2008, BEI Resources released the organism panel (NR-9545) composed of representatives of the diarrheagenic E. coli pathotypes ETEC, EHEC, EPEC, EIEC and EAEC available for ordering. Genomic DNA panels from these organisms are also now available (NR-9546).

Supporting documentation includes strain propagation information and growth and morphology on certain selective and differential microbiological media (see figure to the right).

For technical questions about these products or to learn more, please contact our Technical Services Department.

M. tuberculosis knockout clone pools and companion DeADMAn microarray are now available

Dr. William Bishai , Dr. Gyanu Lamichhane, and colleagues from the Tuberculosis Animal Research and Gene Evaluation Taskforce (TARGET), supported by NIH/NIAID contract N01-AI30036 awarded to the Johns Hopkins University, have generated twenty M. tuberculosis knockout clone pools to facilitate analyses of gene function under in vivo conditions of interest to researchers. In addition, they collaborated with Dr. Robert Fleischmann and Dr. Scott Peterson and colleagues from the Pathogen Functional Genomics Resource Center, supported by NIH/NIAID contract N01-AI15447 awarded to the J. Craig Venter Institute, to develop the DeADMAn Microarray Version 1.0 as a complimentary reagent for use with the knockout clone pools.

One or more M. tuberculosis knockout clone pools containing genetically defined transposon mutants of interest (input pool) are used to infect an animal that is subsequently subjected to a stress condition. Mutants from the input pool and those recovered from the stressed animals (output pool) are identified by amplification of unique transposon junction sequences of each mutant. The competitive hybridization of the input and output pools to the DeADMAn microarray allows for the easy detection of mutants in the input pool and missing from the output pool. The 20 M. tuberculosis knockout clone pools (NR-15773 to NR-15792) and the DeADMAn Microarray Version 1.0 (NR-18958) are now available from BEI.

New Monkeypox Virus Real-Time PCR Assay Discriminates Monkeypox Viruses from other Orthopox Viruses

Monkeypox virus is transmissible to people and it has become the most important Orthopoxvirus infection in human beings since the eradication of smallpox in 1970s.

A Monkeypox Virus quantitative PCR assay (Figure 1) which preferentially detects Monkeypox viruses is now available through BEI Resources (NR-9351). This assay detects Monkeypox viruses at least 1000 fold more efficiently over most other Orthopox viruses or non-Orthopox viruses (Figure 2). Since Monkeypox virus and Vaccinia share considerable homology it is a critical advantage that this assay can detect Monkeypox strains at a minimum of 100 fold more efficiently over the other Vaccinia strains.

The qPCR Monkeypox assay includes the following components: primers, FAM labeled probe, and plasmid for a standard curve. Each vial of reagent can be used for approximately 96 reactions. For technical questions about this assay or to learn more, please contact our Technical Services Department.

Recombinant B. anthracis spore proteins and polyclonal antibodies are now available

Bacillus anthracis are gram positive, spore-forming bacteria. The B. anthracis spore is covered with a balloon-like exosporium that is made up of multiple proteins including the immunodominant glycoprotein Bcl-A (1, 2). Current vaccine strategies have targeted the protective antigen (PA) protein which is an essential component of both lethal toxin and edema toxin. However, vaccines based on immunization with PA alone can result in varied levels of protection in the host (3). More recent research has suggested that an improved protective effect can be achieved by immunizing animals with a combination of PA and other proteins present in the exosporium when compared to immunization with PA alone (1, 4).

BEI Resources has just released recombinant exosporium proteins along with their complementary rabbit polyclonal antibodies. Each protein has been shown experimentally to react with the anti-spore polyclonal antibody 31101-01 (1). In addition, immunization of mice with the hypothetical protein p5303 or the structural protein BxpB in combination with PA showed enhanced protection against spore challenge (1).

Localization of B. anthracis spore proteins within the spore by immunoelectron microscopy. Spores were labeled, following embedding in 4% formaldehyde, by incubation with A) NR-10436 anti-GerQ polyclonal IgG, or B) anti-p5303 polyclonal IgG followed by a gold-labeled secondary antibody (1).

For technical questions about these products, or to learn more, please contact us by clicking here.

Recombinant Protein	Polyclonal Antibody
NR-9577 Collagen-like protein BclA	NR-9578 Polyclonal antibody to BclA
NR-10435 BA5641 Spore coat protein GerQ	NR-10436 Polyclonal antibody to GerQ
NR-10505 BA4499 Superoxide dismutase SODA1	NR-10506 Polyclonal antibody to SODA1
NR-12128 BA1489 Superoxide dismutase SOD15	NR-12129 Polyclonal antibody to SOD15
NR-12130 BA5699 Hypothetical protein p5303	NR-12131 Polyclonal antibody to p5303
NR-12132 BA1237 Hypothetical exosporium protein BxpB	NR-12133 Polyclonal antibody to BxpB

References:

1. Cybulski, R.J., Sanz, P., McDaniel, D., Darnell, S., Bull, R.L., and O’Brien, A.D. Recombinant Bacillus anthracis spore proteins enhance protection of mice primed with suboptimal amounts of protective antigen. Vaccine 2008;26:4927-4939.

2. Sylvestre, P., Couture-Tosi, E., and Mock, M. A collagen-like surface glycoprotein is a structural component of the Bacillus anthracis exosporium. Mol Microbiol 2002;45(July(1)):5240-7.

3. Fellows, P.F., Linscott, M.K., Ivins, B.E., Pitt, M.L., Rossi, C.A., Gibbs, P.H. et al. Efficacy of a human anthrax vaccine in guinea pigs, rabbits and rhesus macaques against challenge by Bacillus anthracis isolates of diverse geographical origin. Vaccine 2001:19(April(23-24)):3241-7.

4. Brahmbhatt, T.N., Darnell, S.C., Carvalho, H.M., Sanz, P., Kang T.J., Bull, R. L., et. al. Recombinant exosporium protein BclA of Bacillus anthracis is effective as a booster for mice primed with suboptimal amounts of protective antigen. Infect Immun 2007;75 (November (11)):5240-7.

Ferret Reagents available through BEI Resources

In recent years, the ferret has proven to be an excellent animal model for the study of pathogenicity and transmissibility of influenza viruses (Journal of Virology 81 (13): 6890-9898 (July 2007)). Even the attachment of highly pathogenic avian influenza H5N1 in the lower respiratory track of humans is mimicked in the ferret model (Science 312 (5772): 5772 (April 21, 2006)). In like manner, ferrets are also being shown to be a good model for SARS research because they are susceptible to infection by the SARS coronavirus and can transmit the virus to uninfected animals (Nature 425: 915 (October 30, 2003)).

To support the research on these emerging infections, BEI Resources (under the direction of NIAID) will soon offer a vast array of tools for the use of the ferret model. In February of 2008, BEI Resources made available approximately 100 ferret immune gene primers that were produced under contract by the laboratory of Dr. David Kelvin (Head, Division of Experimental Therapeutics, University Health Network). In Spring 2008, BEI Resources will also offer a number of monoclonal antibodies against ferret immune antigens, also from Dr. Kelvin’s laboratory.

Immortalized murine macrophages and microglial cell lines from TLR knockout mice available

The Toll-like receptor (TLR) family is the essential recognition and signaling component of multicellular organisms against all the major classes of pathogens. The study of TLR signaling pathways has become a productive area of investigation for researchers interested in signal transduction during innate immunity and inflammation. Of interest to biodefense, the identification of TLRs, their ligands, and signaling events will provide insight into understanding the role of the innate immune response in infectious diseases such as anthrax, plague, tularemia, and smallpox. Knockout mice for many of these innate immune receptors and adaptor molecules are not always widely available. In addition, generation of such knockout animals requires significant human and economic resources.

BEI Resources houses a number of immortalized macrophage cell lines from knockout mice deficient in TLRs and their signaling molecules. Characterization of the immortalized cell lines has shown a deficiency in the production of cytokines upon stimulation with defined differential TLR ligands using ELISA (Fig. 1).

Use of this resource will enable the investigation of immune mechanisms against pathogens and evaluation of pharmacological agents. For a list of available cell lines or technical questions about these products, please contact us by clicking here.

Fig. 1. Secretion profiles of TNF among immortalized cell lines derived from knockout mice lacking the TLR adaptor molecules MAL/TIRAP and MyD88. Cells were stimulated with increasing concentrations of ligands against TLR2/6 (Pam2), TLR3 (PolyIC), and TLR4 (LPS). A, cytokine secretion in wild type and MAL/TIRAP -/- macrophages. B, cytokine secretion in wild type and MyD88 -/- microglial cells.

Epsilon Toxin Reagents are now Available from BEI Resources

Clostridium perfringens are gram positive, obligate anaerobes that produce multiple toxins. Type B and D strains, responsible for necrotic enteritis or enterotoxemia in animals, express Epsilon toxin encoded by the etx gene. This protein exerts its cytotoxic activity on sensitive cells by forming pores in the cell membrane resulting in a loss of cellular homeostasis. Epsilon toxin is a USDA and HHS select agent toxin and is included on the NIAIDlist of Category B priority pathogens.

BEI Resources has reagents that can be used to study the molecular mechanisms of Epsilon toxin cytotoxicity. NR-856 is native Epsilon protoxin purified directly from C. perfringens culture supernatant. NR-4670, the native protoxin that has been activated using trypsin, is cytotoxic to ATCC® CCL-34™ MDCK cells (Figure B). In addition, we offer NR-865 polyclonal antibody to Epsilon toxin which can detect the protein in an ELISA or western blot format.

Fluorescence micrographs showing MDCK cells treated with NR-4670 activated Epsilon toxin (B) or without toxin (A) for 1.5 hours in the presence of propidium iodide. Pore formation by the toxin is demonstrated by the uptake and fluorescence of propidium iodide compared to the untreated control.

For technical questions about these products, or to learn more, please contact us by clicking here.

Yersinia pestis Plasmid Profiling Assay is Now Available

The agent of bubonic plague Yersinia pestisis a NIAID Category A Priority Pathogen and likely biothreat. In support of research and product development in this area BEI Resources released a Yersinia pestis Plasmid Detection Kit (NR-9562) in April 2008.

This assay allows the researcher to quickly identify the presence of plasmid-encoded virulence factors pPCP1, pMT1 and pCD1 and is designed to be used for characterization purposes in conjunction with standard microbiological techniques.

The assay consists of primer sets designed to specifically detect the pPCP1, pMT1 and pCD1 plasmids using standard polymerase chain reactions and includes a species-specific positive control primer set and internal positive control template.

PCR amplification of genomic DNA from Yersinia pestis displaying a pPCP1+, pMT1+, pCD1– profile (lanes 2-4 respectively). Lanes 5 and 6 are the Yersinia pestis and internal positive controls. Lane 7 is a non-template control."

Panel A. Growth of Yersinia pestis on Congo Red media showing red/orange colonies typical of pgm+ organisms. Organisms that are confirmed pgm+ and pCD1+ are considered Select Agents.

Panel B. Bacteria lacking pgm showing distinctive variations in colony morphology and reduced coloring on Congo Red media as compared to pgm+ organisms.

For technical questions about these products or to learn more, please contact our Technical Services Department.

BEI Resources offers Overlapping Peptide Arrays for Pathogens

Custom peptides have many uses in biotechnology including epitope mapping, analyzing protein-protein interactions, and as antigens for the generation of anti-peptide antibodies. For epitope mapping, arrays consisting of a small amount of a large number of overlapping peptides are required. For the individual researcher, the generation of such a custom array of overlapping peptides for specific proteins of interest could be cost-prohibitive. For that reason, NIAID has funded BEI Resources to generate a substantial number of peptide arrays related to applications in biodefense and emerging infections. These reagents are provided free of charge (except for shipping and handling fees) for use by BEI registrants.

Currently BEI Resources offers peptide arrays for proteins of influenza, SARS-CoV and human coronaviruses, dengue virus, hepatitis C virus, West Nile virus, hantavirus, vaccinia virus, Yersinia pestis, and Bacillus anthracis (complete list). For more information on these peptide arrays, please search our online catalog for your peptide array of interest. If you need more assistance or information about BEI Resources peptide arrays, please contact us by clicking here.

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