DNA FINGERPRINTING OF MYCOBACTERIUM BOVIS AN5 STRAIN BY RFLP TECNIQUE

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HEIDARZADE M.; SALEHI M.; AREF PAJOHO R.; BOLFION M.; SOLYMANI K.; MOSAVARI N.

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Journal: JOURNAL OF MICROBIOLOGY KNOWLEDGE Year:2009 | Volume:1 | Issue:4 Page(s): 37-44

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Title

DNA FINGERPRINTING OF MYCOBACTERIUM BOVIS AN5 STRAIN BY RFLP TECNIQUE

Author(s)

Keywords

MYCOBACTERIUM BOVIS AN5 STRAINQ4

BOVINE TUBERCLOSISQ4

TUBERCULINQ4

RFLPQ4

HYBRIDATIONQ4

DR & PGRS PROBESQ4

Abstract

Mycobacterium bovis in comparision to other members of Mycobacterium tuberclusis complex has largest host capacity. In veterinary medicine TUBERCULINation and killing the positive reactors remains one of the most acceptable strategy in control of bovine tuberclusis in bovids. RAZI Institute(RVSRI) is the exclusive national supplier of TUBERCULIN from since 1940 until now in IRAN anf since 2004 until now that bovine TUBERCULIN supersending mammalian TUBERCULIN for diagnosis bovine tuberclusis, MYCOBACTERIUM BOVIS AN5 STRAIN superseding Mycobacterium toberclusis strains for production bovine TUBERCULIN.In result of consecutive and compulsory subcultures of the strain for produce this product genetic stability of this exotic strain, the sub_culture’s of this strain has experienced to ensure from non_exictence genetic variation. To determine<this investigation is for consideration genetic variation in consecutive sub_cultures of MYCOBACTERIUM BOVIS AN5 STRAIN.At first, for proving be Mycobacterium bovis, our strain in this consideration, one piece of likeness gen oxyR in length 548 bp considered in this research, nucleotide of 285 of this piece in genom of Mycobacterium bovis has adenine but in all of the members of Mycobacterium tuberclusis has guanine and couse to disappear a part of revenue Alu I enzyme. So, in this piece on Mycobacterium bovis is 4 cut’s part and in other members of Mycobacterium tuberclusis complex is 3 cut’s part for this enzyme. In AN5 strain, the piece of likeness gen oxyR multiply with PCR technique then cut with AluI enzyme and considered cut’s pattern. Then In order to ensure that the currently available M. bovis AN5 strain at the institute has not experienced major genetic events, five consecutive fresh sub-cultures of the strain on plain Lowenstein-Jensen (L J) and pyruvate-supplemented L J culture media were subjected to bacterial DNA extraction. These DNA templates were subsequently digested by PvuII and genotyped by PGRS-RFLP and DR-RFLP. Result of first consider with likeness gen oxyR and anzymatic digestion with AluI show that our template is mycobacterium bovis. In consider of subcultures, result of enzymatic digestion with PvuII and AluI and HYBRIDATION with DR probe, both of them shown presence of three DR’S region in all of considerable subcultures that 3 bands of enzymatic digestion with AluI and HYBRIDATION with DR probes in all of considerable subcultures was downer from 2027bp and 3 bands of enzymatic digestion with PvuII and HYBRIDATION with DR probes in all of considerable subcultures were middle of 4371bp and 2322bp and a band was in downer of 2027bp. In consider the result of enzymatic digestion with AluI and PvuII and hybrydation with PGRS probe, observation many bands that number of bands and molecular sizes in all of subcultures were identical and no difference obtained.Scrutinizing, an identical in molecular size and the number of bands in patterns obtained with DR&PGRS_RFLP technique, show non_exictence genetic variation in 5 subcultures of this strain on plain Lowenstein_jensen and pyruvate.

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References

MOLECULAR DIFFERENTIATION BETWEEN BOVINE MYCOBACTERIUM BOVIS ISOLATES AND HUMAN MYCOBACTERIUM TUBERCULOSIS ISOLATES IN IRAN THROUGH PCR-RFLP METHOD AND COMPARISON WITH 5 STANDARD STRAINS

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APA: Copy

HEIDARZADE, M., SALEHI, M., AREF PAJOHO, R., BOLFION, M., SOLYMANI, K., & MOSAVARI, N.. (2009). DNA FINGERPRINTING OF MYCOBACTERIUM BOVIS AN5 STRAIN BY RFLP TECNIQUE. JOURNAL OF MICROBIOLOGY KNOWLEDGE, 1(4), 37-44. SID. https://sid.ir/paper/193240/en

Vancouver: Copy

HEIDARZADE M., SALEHI M., AREF PAJOHO R., BOLFION M., SOLYMANI K., MOSAVARI N.. DNA FINGERPRINTING OF MYCOBACTERIUM BOVIS AN5 STRAIN BY RFLP TECNIQUE. JOURNAL OF MICROBIOLOGY KNOWLEDGE[Internet]. 2009;1(4):37-44. Available from: https://sid.ir/paper/193240/en

IEEE: Copy

M. HEIDARZADE, M. SALEHI, R. AREF PAJOHO, M. BOLFION, K. SOLYMANI, and N. MOSAVARI, “DNA FINGERPRINTING OF MYCOBACTERIUM BOVIS AN5 STRAIN BY RFLP TECNIQUE,” JOURNAL OF MICROBIOLOGY KNOWLEDGE, vol. 1, no. 4, pp. 37–44, 2009, [Online]. Available: https://sid.ir/paper/193240/en

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