Giardia intestinalis (Lambl) Alexeieff
(ATCC? 50581?) 統(tǒng)一編號(hào)
Strain Designations 別名: GS clone H7 /
Depositor: TE Nash /
Biosafety Level 生物安全等級(jí) : 2
Application Enteric Research Food and waterborne pathogen research
Biosafety Level 生物安全等級(jí) 2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. Isolation Unknown
Product Format 提供形式 frozen
Storage Conditions 保藏方法
Frozen Cultures: -70℃ for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures: 2-8℃ Live
Cultures: See Protocols section for handling information
Type Strain no
Genome Sequenced Strain Yes
Comments注釋 Cysteine-rich variant surface proteins
Frequency of variant antigens
Antigenic variation
Purification of variant-specific surface protein
Epitope variability in susceptibility to proteases
Metabolism of cysteine
Association of variant surface antigens with thick cell coat
Differentiation induced by cholesterol starvation
Variant specific epitopes
Control of infections in mice
Genome sequencing strain
Medium 培養(yǎng)基 ATCC? Medium 2695: Keister's Modified TYI-S-33
ATCC? Medium 2155: LYI Giardia Medium (filtered)
Growth Conditions 生長條件
Temperature 培養(yǎng)溫度: 35℃
Atmosphere 需氧情況: Anaerobic
Culture System 文化系統(tǒng): Axenic
Cryopreservation 1，Harvest and Preservation Harvest cells from a culture that is at or near peak density. To detach cells from the wall of the culture tubes place on ice for 10 minutes. Invert tubes several times until the majority of the cells are in suspension. Centrifuge tubes at 800 x g for 5 minutes,
2， Adjust the concentration of cells to 2 x 107/mL in fresh medium.
3，Before centrifuging prepare a 24% (v/v) solution of sterile DMSO in fresh medium containing 8% (w/v) sucrose. The solution is prepared as follows:
a，Add 1.05 g sucrose to 10 mL of fresh medium and filter sterilize through a 0.2 ?m filter;
b，Add 2.4 mL of DMSO to an ice cold 20 x 150 mm screw-capped test tube;
c， Place the tube on ice and allow the DMSO to solidify (~5 min) and then add 7.6 mL of ice cold medium prepared in step 3a. The final concentration will be 24% (v/v) DMSO and 8% (w/v) sucrose;
d， Invert several times to dissolve the DMSO;
e，Allow to warm to room temperature.
4，Mix the cell preparation and the cryoprotective agent, prepared in step 3, in equal portions. Thus, the final concentration will equal 12% (v/v) DMSO + 4% sucrose (w/v) and 107 cells/mL. The time from the mixing of the cell preparation and DMSO stock solution to the start of the freezing process should be no less than 15 min and no longer than 30 min.
5，Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation). Place the vials in a controlled rate freezing unit. From room temperature cool at -1℃/min to -40℃. If the freezing unit can compensate for the heat of fusion, maintain rate at -1℃/min through the heat of fusion. At -40℃ plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1℃ freezing apparatus.
6，Place the apparatus at -80℃ for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1℃/min.)
7，The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130℃ are stabile indefinitely. Those stored at temperatures above -130℃ are progressively less stabile as the storage temperature is elevated.
8， To establish a culture from the frozen state place an ampule in a water bath set at 35℃. Immerse the vial just to a level just above the surface of the frozen material. Do not agitate the vial.
9，Immediately after thawing, do not leave in the water bath, aseptically remove the contents of the ampule and inoculate a 16 x 125 mm screw-capped test tube containing 13 mL ATCC Medium 2695. Incubate the culture on a 15? horizontal slant at 35℃.
Name of Depositor 寄存人TE Nash Special Collection NCRR Contract
References 參考文獻(xiàn) Aggarwal A, et al. Cysteine-rich variant surface proteins of Giardia lamblia. Mol. Biochem. Parasitol. 32: 39-48, 1989. PubMed: 2911277
Elmendorf HG, et al. The abundance of sterile transcripts in Giardia lamblia. Nucleic Acids Res. 29: 4674-4683, 2001. PubMed: 11713317
Gottstein B, et al. Antigenic variation in Giardia lamblia: cellular and humoral immune response in a mouse model. Parasite Immunol. 12: 659-673, 1990. PubMed: 1707508 Gottstein B, Nash TE. Antigenic variation in Giardia lamblia: infection of congenitally athymic nude and scid mice. Parasite Immunol. 13: 649-659, 1991. PubMed: 1725820
Gottstein B, et al. Human cellular immune response to Giardia lamblia. Infection 19: 421-426, 1991. PubMed: 1816113
Langford TD, et al. Central importance of immunoglobulin A in host defense against Giardia spp. Infect. Immun. 70: 11-18, 2002. PubMed: 11748158
Lujan HD, et al. Purification of a variant-specific surface protein of Giardia lamblia and characterization of its metal-binding properties. J. Biol. Chem. 270: 13807-13813, 1995. PubMed: 7775437
Lujan HD, et al. Cholesterol starvation induces differentiation of the intestinal parasite Giardia lamblia. Proc. Natl. Acad. Sci. USA 93: 7628-7633, 1996. PubMed: 8755526
Lujan HD, Nash TE. The uptake and metabolism of cysteine by Giardia lamblia trophozoites. J. Eukaryot. Microbiol. 41: 169-175, 1994. PubMed: 8167619
Marti M, et al. Conformationally correct expression of membrane-anchored Toxoplasma gondii SAG1 in the primitive protozoan Giardia duodenalis. Infect. Immun. 70: 1014-1016, 2002. PubMed: 11796643
Murtagh JJ Jr., et al. Guanine nucleotide-binding proteins in the intestinal parasite Giardia lamblia. Isolation of a gene encoding an approximately 20-kDa ADP-ribosylation factor. J. Biol. Chem. 267: 9654-9662, 1992. PubMed: 1577802
Nash TE, et al. Antigenic variation of Giardia lamblia in experimental human infections. J. Immunol. 144: 4362-4369, 1990. PubMed: 2341723
Nash T, Rice WG. Efficacies of zinc-finger-active drugs against Giardia lamblia. Antimicrob. Agents Chemother. 42: 1488-1492, 1998. PubMed: 9624499
Nash TE, et al. Frequency of variant antigens in Giardia lamblia. Exp. Parasitol. 71: 415-421, 1990. PubMed: 1699782
Nash TE, et al. Isolate and epitope variability in susceptibility of Giardia lamblia to intestinal proteases. Infect. Immun. 59: 1334-1340, 1991. PubMed: 1706319
Nash TE, et al. Variant specific epitopes of Giardia lamblia. Mol. Biochem. Parasitol. 42: 125-132, 1990. PubMed: 1700296
Pimenta PF, et al. Variant surface antigens of Giardia lamblia are associated with the presence of a thick cell coat: Thin section and label fracture immunocytochemistry survey. Infect. Immun. 59: 3989-3996, 1991. PubMed: 1937758
Singer SM, et al. Biological selection of variant-specific surface proteins in Giardia lamblia. J. Infect. Dis. 183: 119-124, 2001. PubMed: 11087204
Singer SM, Nash TE. T-cell-dependent control of acute Giardia lamblia infections in mice. Infect. Immun. 68: 170-175, 2000. PubMed: 10603384
Touz MC, et al. The activity of a developmentally regulated cysteine proteinase is required for cyst wall formation in the primitive eukaryote Giardia lamblia. J. Biol. Chem. 277: 8474-8481, 2002. PubMed: 11773053
Cross References
Nucleotide (GenBank) : ACGJ01000000 Giardia intestinalis ATCC 50581 strain GS/M clone H7, whole genome shotgun sequencing project.
