CLONING AND PARTIAL SEQUENCE OF TRANSFERRIN-BINDING PROTEIN-2 OF NEISSERIA-MENINGITIDIS USING A NOVEL METHOD - TWIN N-TERMINAL PCR

被引：4

作者：

WILTON, J ^{[1
]}

ALAALDEEN, D ^{[1
]}

PALMER, HM ^{[1
]}

BORRIELLO, SP ^{[1
]}

机构：

[1] CLIN RES CTR, MICROBIAL PATHOGEN RES GRP, HARROW HA1 3UJ, MIDDX, ENGLAND

来源：

FEMS MICROBIOLOGY LETTERS | 1993年 / 107卷 / 01期

关键词：

NEISSERIA-MENINGITIDIS; NEISSERIA-GONORRHOEAE; TRANSFERRIN; POLYMERASE CHAIN REACTION; TWIN N-TERMINAL POLYMERASE CHAIN REACTION;

D O I：

10.1111/j.1574-6968.1993.tb06004.x

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The genes encoding transferrin-binding proteins (TBPs) 1 and 2 of Neisseria meningitidis and N. gonorrhoeae were used as model loci in a novel method of cloning (twin N-terminal polymerase chain reaction; TNT-PCR) involving amplification between the 5' ends of two genes. Primers were based on N-terminal amino-acid sequences. A 2.1-kb product amplified from N. meningitidis strain SD (B15 P1.16) was cloned into a plasmid vector and partially sequenced. Translated sequence immediately downstream of the primer at both ends of this product correlated to the additional known N-terminal amino acids of TBP-1 and 2. The protein encoded by the cloned sequence reacted with TBP-2-specific antiserum. The size of products generated in TNT-PCR correlated exactly with the different sized TBP-2 produced by 10 strains of the Neisseria spp. examined, indicating successful cloning of the gene for TBP-2 and showing it to be adjacent to and preceding TBP-1 on the chromosome for both N. meningitidis and N. gonorrhoeae.

引用

页码：59 / 66

页数：8

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