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KMID : 0545120030130020287
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 2 p.287 ~ p.291
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Effects of Minor Arginyl tRNA and Isoleucyl tRNA on the Expression of Clostridium botulinum Neurotoxin Light Chain in Escherichia coli
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KIM, JIN SOOK
SEONG, HYE YOUNG/KIM, MI WHA/KU, JONG SEO/CHOI, SOON-YONG
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Abstract
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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 if recombinant protein. The argD and ileX tRNA genes were cloned into pACTC184 vector, resulting in pAAD13IX plasmid. The translational stress of the toxin gene related to codon bias was reversed by supplementation of the AGA arginyl tRNA of T4 phage and AUA isoleucyl tRNA of E, coli. This system may be applicable for expression of a variety of AT-rich heterologous genes in E. coli.
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