A reliable amplification technique for the characterization of genomic DNA sequences flanking insertion sequences

被引：61

作者：

Prod'hom, G

Lagier, B

Pelicic, V

Hance, AJ

Gicquel, B

Guilhot, C

机构：

[1] Inst Pasteur, Unite Genet Mycobacterienne, F-75724 Paris 15, France

[2] Inst Pasteur, Ctr Nacl Reference Mycobacteries, F-75724 Paris, France

[3] Fac Xavier Bichat, INSERM, U82, Paris, France

来源：

FEMS MICROBIOLOGY LETTERS | 1998年 / 158卷 / 01期

关键词：

Mycobacterium; cloning; insertion sequence; transposon mutagenesis;

D O I：

10.1111/j.1574-6968.1998.tb12803.x

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A simple and efficient ligation-mediated PCR (LMPCR) is described for amplifying DNA adjacent to known sequences. The method uses one primer specific for the known sequence and a second specific for a synthetic linker ligated to restricted genomic DNA. Perkin-Elmer AmpliTaq Gold polymerase is used to minimize non-specific primer annealing and amplification. This LMPCR method was successfully applied to isolate DNA sequences flanking mobile elements present in mycobacterial mutants generated by transposon mutagenesis. (C) 1998 Published by Elsevier Science B.V. All rights reserved.

引用

页码：75 / 81

页数：7

共 11 条

[1]

BUTTLER WR, 1996, J CLIN MICROBIOL, V34, P1801

[2] PREVENTION OF PRE-PCR MIS-PRIMING AND PRIMER DIMERIZATION IMPROVES LOW-COPY-NUMBER AMPLIFICATIONS [J].