酿酒酵母中表达神经肉毒素A及其功能研究

Expression of Botulinum Neurotoxins A inSaccharomyces cerevisiaeand Its Functional Research

DOI：10.3969/j.issn.1673-1689.2019.02.013

中文关键词: 肉毒神经毒素A（BoNT/A） 酿酒酵母 SEC9

英文关键词: botulinum neurotoxins A（BoNT/A）,Saccharomyces cerevisiae,SEC9

基金项目:

摘要点击次数: 296

全文下载次数: 282

中文摘要:

肉毒神经毒素（BoNT）是肉毒梭菌产生的神经毒素，是最强大的毒素之一。为了研究肉毒神经毒素A在酵母中的活性及功能，在酿酒酵母细胞中表达 BoNT/A LC，观察其对酵母生长影响以及通过蛋白免疫印迹方法研究其在酵母中蛋白表达水平。结果表明：BoNT/A抑制野生型酵母的生长；在酿酒酵母细胞中BoNT/A具有水解酶活性，当其活性位点突变后，BoNT/A突变体的表达不会抑制酿酒酵母的生长；过量表达酵母的SNAP25同源基因SEC9可以回补BoNT/A轻链过表达的酵母的生长。在BoNT/A轻链表达的酵母细胞中，其催化活性可以通过酵母的生长来检测。因此，这类特殊的酵母细胞可以用来分析BoNT/A。

英文摘要:

Botulinum toxin（BoNT） is a neurotoxin produced byClostridium botulinum，which is one of the most powerful toxin. To study the activity and function of BoNT/A in yeast，BoNT/A LC in yeast is expressed，observing the yeast growth and BoNT/A protein expressing level by western blot. Expression of LC of BoNT cerotype A caused a growth defect in yeast cells，and the toxicity is due to the hydrolytic activity. It is because catalytically inactive BoNT/A LC does not cause a growth defect in yeast cells. Overexpression of a yeast SNAP-25 homologue gene，SEC9，could rescue the growth defect in BoNT/A LC expressing cells. The results demonstrate that，in BoNT/A LC expressing yeast cells，the hydrolytic activity can be monitored through their growth. Thus，such cells can be used to analyze BoNT/A.

查看全文 查看/发表评论 下载PDF阅读器