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目的克隆编码虎纹捕鸟蛛多肽的基因并表达蜘蛛多肽,为研究其生物学功能奠定基础。方法通过构建和随机测序毒腺cDNA文库,克隆编码毒素多肽的基因;构建真核表达质粒,利用酿酒酵母S78株表达系统分泌表达,表达产物进行Tricine SDS-PAGE鉴定。结果克隆到一种编码富含半胱氨酸的多肽的新基因,命名为HWTX-XIVa2基因;构建的真核表达质粒转化酵母S78后表达6.5ku的多肽。结论构建并筛选毒腺cDNA文库是发现毒素多肽基因的一种有效方法,利用基因工程表达是获得蜘蛛多肽的一种有效手段。
OBJECTIVE: To clone the gene encoding the polypeptide of the spiders and express the spider polypeptide, and lay the foundation for the study of its biological functions. Methods The cDNA encoding the toxin polypeptide was constructed and sequenced randomly. The eukaryotic expression plasmid was constructed and secreted by Saccharomyces cerevisiae S78 strain. The expressed product was identified by Tricine SDS-PAGE. Results A new gene encoding cysteine-rich polypeptide was cloned and named as HWTX-XIVa2. The constructed eukaryotic expression plasmid transformed yeast S78 to express 6.5ku polypeptide. Conclusion The construction and screening of the cDNA library of the glandular gland is an effective method for the discovery of the toxin polypeptide gene. The use of genetic engineering expression is an effective means of obtaining the spider polypeptide.