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Soluble expression of the antifungal peptide genes, Drs and Drs-lC, of Drosophila melanogaster in the prokaryote and the detection of their antifungal activity
Author(s): 
Pages: 747-753
Year: Issue:  5
Journal: ACTA ENTOMOLOGICA SINICA

Keyword:  黑腹果蝇抗真菌肽原核表达可溶性表达抗真菌活性;
Abstract: Drosomycin(Drs)是第1个从黑腹果蝇Drosophila melanogaster体内鉴定发现的昆虫抗真菌肽因子.它对细菌无明显的抗性,但对丝状真菌具有高效广谱的抑杀作用.此外,在黑腹果蝇基因组还存在着Drs的另外6个同系物的基因序列,其中同系物Drosomycin-lC(Drs-lC)的抗真菌谱仅次于Drs.将Drs抗真菌肽基因(Drs)和同系物Drs-lC基因(Drs-lC)进行可溶性表达,对果蔬等农产品防腐保鲜的研究有应用前景.本实验将Drs和Drs-lC分别克隆到硫氧还蛋白(Trx)融合表达载体pThiohis A中,转化宿主菌TOP10,进行可溶性表达,并从诱导表达的菌液起始浓度、IPTG的诱导浓度及诱导时间等方面进行了表达条件的优化.结果表明2种融合蛋白Trx-Drs和Trx-Drs-lC大部分以可溶形式表达,可溶性表达的Trx-Drs在上清液中约占菌体总蛋白的22%.2种融合蛋白的表达产物经Ni-NTA亲和层析得到纯化.生测结果表明,2种融合蛋白分别对8种供试真菌中的5种真菌显示明显的抗性.
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