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Cloning, sequencing and expression of the transferrin-binding protein 2 gene from Actinobacillus pleuropneumoniae

Published online by Cambridge University Press:  12 February 2007

Wang Fang
Affiliation:
Key Laboratory of Animal and Poultry Diseases Diagnostic, Ministry of Agriculture; Institute of Veterinary Medicine of Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
He Kong-Wang*
Affiliation:
Key Laboratory of Animal and Poultry Diseases Diagnostic, Ministry of Agriculture; Institute of Veterinary Medicine of Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
*
*Corresponding author: Email kwh2003@263.net

Abstract

To amplify the transferrin-binding protein 2 (Tbp2) gene from Actinobacillus pleuropneumoniae (App) by polymerase chain reaction (PCR), a pair of primers were designed according to sequence Z46774 of A. pleuropneumoniae serotype 5. The amplified DNA fragment (1642 bp) was cloned into pMD18-T and sequenced. The sequencing result showed that the homology was 99.8% compared with the reference sequence. The target fragment of the positive clones was inserted into pET-32a and transformed in Escherichia coli BL21 (DE3). After its induction, the fragment was expressed in E. coli. The Western blotting results were positive.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

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