[KSCI] Korea Science Citation Index Service

Effects of Minor Arginyl tRNA and Isoleucyl tRNA on the Expression of Clostridium botulinum Neurotoxin Light Chain in Escherichia coli

Kim, Jin-Sook (Department of Microbiology, HanNam University)
Seong, Hye-Young (Department of Microbiology, HanNam University)
Kim, Mi-Wha (Department of Microbiology, HanNam University)
Ku, Jong-Seo (Department of Microbiology, HanNam University)
Choi, Soon-Yong (Department of Microbiology, HanNam University)

Publication Information

Journal of Microbiology and Biotechnology / v.13, no.2, 2003 , pp. 287-291 More about this Journal

Abstract

Botulinum neurotoxin type A (BONT/A) is an extremely potent toxin, which is produced by Clostridium botulinum. The light chain of this protein (BONT/A LC), which is known as a zinc endopeptidase, cleaves SNAP-25 involved in the exocytosis process. In this work, the expression of recombinant BoNT/A LC in E. coli is described. The BONT/A LC gene of C. botulinum contains a high frequency of the arginine AGA and isoleucine ATA codons that are rarely used in genes of E. coli, hampering the translation of recombinant protein. The argD and ilex tRNA genes were cloned into pACYC184 vector, resulting in pAAD131X plasmid. The translational stress of the toxin gene related to codon bias was reversed by fupplernentation of the AGA arginyl tRNA of T4 phage and AUA isoleucyl tRNA of E. coli. This system may be applicable for the expression of a variety of AT-rich heterologous genes in E. coli.

Keywords

Neurotoxin; codon-bias; tRNA; Clostridium botulinum;

Citations & Related Records

Times Cited By KSCI : 4 (Citation Analysis)
Times Cited By Web Of Science : 7 (Related Records In Web of Science)

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