Cloning of Clostridium Perfringens Iota Toxin Gene in Escherichia Coli

AUTHORS

P. Seyed Sayyah 1 , B. Golestani 1 , * , R. Pilehchian Langroud 2

AUTHORS INFORMATION

1 Department of Biology, Islamic Azad University, Urmia branch, Urmia, Iran

2 Specialized Clostridia reseach laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Alborz, Karaj, Iran

ARTICLE INFORMATION

Archives of Razi Institute: 73 (2); 107-111
Published Online: March 01, 2018
Article Type: Journal Article
Received: January 14, 2017
Accepted: April 26, 2017
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Abstract

Iota toxin is produced by Clostridium perfringens type E. This toxin causes antibiotic-associated enterotoxemia in lambs and calves. Iota toxin is a binary toxin that has two components including Ia (the enzyme component) and Ib (the binding component). Ib binds to the surface receptor of target cells and translocate Ia into the cytosol of cells. The aim of this study was to clone toxigenic epitope of iota a gene in E. coli strain Top10. In this study, the phenol–chloroform method was used for the extraction of the whole genomic DNA. The toxigenic epitope of iota a gene was amplified by polymerase chain reaction (PCR). The PCR product was ligated into the pTZ57R/T vector cloning site. Then, based on the TA-cloning method, the product was cloned in competent E. coli strain Top10. Colony PCR was used to screen bacterial colonies transformed with recombinant plasmids. The presence of 446-bp fragment on agarose gel showed that the toxigenic epitope of iota a gene of C. perfringens has been cloned in E. coli strain Top10.

Keywords

Cloning Clostridium perfringens Iota toxin E. coli TA-cloning

© 2018, Archives of Razi Institute. Razi Vaccine and Serum Research Institute.
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