CREATION OF A NULL MUTATION IN THE STX2 GENE OF ESCHERICHIACOLI O157:H7

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FIROUZI R.

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Journal: Journal of Veterinary Research Year:2006 | Volume:61 | Issue:4 Page(s): 357-361

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Title

CREATION OF A NULL MUTATION IN THE STX2 GENE OF ESCHERICHIACOLI O157:H7

Author(s)

FIROUZI R. | Issue Writer Certificate

Keywords

MUTATIONQ3

ESCHERICHIA COLIQ3

SANS-SERIF">O157:H7Q3

SANS-SERIF"> STX2Q3

SUICIDE VECTORQ3

Abstract

Objective: To construct a specific MUTATION in E.coli O157: H7.Design: Allelic exchange approach for producing stx2-negative mutant.Methods:1- Obtaining PCR product from stx2 gene.2- Cloning this product to T-vector.3- Inserting a gentamicin resistance cassette.4- Transfering the stx2 gene with GMR cassette to SUICIDE VECTOR.5- Mating donor (SM10 (+pRE107-stx2: GM) and recipient (StrREC 960904).6- Selection of single-cross-overs.7- Counter-selection of double cross-overs. Selecting GmR/StrR and Amps colonies.8- Confirmation of mutant by PCR and cytotoxigenicity assay on vero cells.Results: In this study, after mating between the donor (SM10 (+pRE107 - Stx2: GM) and the recipient cells (Str R-EC960904 wild type), colonies were resistant to gentamycin and streptomycin but sensitive to ampicillin .The result of PCR showed that the mutant was the stx2 negative strain. However, the mutant strain was unable to lyse the vero cells compare to wild type strain.Conclusion: Hutant strain was produced by a suitable SUICIDE VECTOR.

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APA: Copy

FIROUZI, R.. (2006). CREATION OF A NULL MUTATION IN THE STX₂ GENE OF ESCHERICHIACOLI O₁₅₇:H₇. JOURNAL OF VETERINARY RESEARCH, 61(4), 357-361. SID. https://sid.ir/paper/34614/en

Vancouver: Copy

FIROUZI R.. CREATION OF A NULL MUTATION IN THE STX₂ GENE OF ESCHERICHIACOLI O₁₅₇:H₇. JOURNAL OF VETERINARY RESEARCH[Internet]. 2006;61(4):357-361. Available from: https://sid.ir/paper/34614/en

IEEE: Copy

R. FIROUZI, “CREATION OF A NULL MUTATION IN THE STX₂ GENE OF ESCHERICHIACOLI O₁₅₇:H₇,” JOURNAL OF VETERINARY RESEARCH, vol. 61, no. 4, pp. 357–361, 2006, [Online]. Available: https://sid.ir/paper/34614/en

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