Construction of an Alpha Toxin Gene Knockout Mutant of Clostridium perfringens Type A by Use of a Mobile Group II Intron-Reference-Cited by-同舟云学术

Construction of an Alpha Toxin Gene Knockout Mutant of Clostridium perfringens Type A by Use of a Mobile Group II Intron

Published:2005-11 Issue:11 Volume:71 Page:7542-7547
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Chen Yue¹,McClane Bruce A.²,Fisher Derek J.²,Rood Julian I.³,Gupta Phalguni¹

Affiliation:

1. Department of Infectious Diseases and Microbiology, Graduate School of Public Health

2. Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania

3. Department of Microbiology, Monash University, Victoria, Australia

Abstract

ABSTRACT In developing Clostridium perfringens as a safe vaccine vector, the alpha toxin gene ( plc ) in the bacterial chromosome must be permanently inactivated. Disrupting genes in C. perfringens by traditional mutagenesis methods is very difficult. Therefore, we developed a new strategy using group II intron-based Target-Tron technology to inactivate the plc gene in C. perfringens ATCC 3624. Western blot analysis showed no production of alpha toxin protein in the culture supernatant of the plc mutant. Advantages of this technology, such as site specificity, relatively high frequency of insertion, and introduction of no antibiotic resistance genes into the chromosome, could facilitate construction of other C. perfringens mutants.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/AEM.71.11.7542-7547.2005

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