在大肠杆菌中表达具有抗黄曲霉活性的鲎素  

Tachyplesin Expressed in E.coli Exhibiting Inhibition Activity against Aspergillus Flavus

作　　者：张春义[1] 范云六[1] 

机构地区：[1]中国农业科学院生物技术研究中心

出　　处：《高技术通讯》1999年第1期43-46,共4页Chinese High Technology Letters

基　　金：国际洛克菲勤基金会

摘　　要：人工设计并合成了抗真菌多肽鲎素基因。将合成的鲎素基因首先克隆到ｐＵＣ１８载体上，利用通用引物测序，获得了与预期碱基序列完全一致的目的基因。再将鲎素基因转入大肠杆菌，诱导表达融合蛋白。３７℃下诱导表达的ＧＳＴ—鲎素融合蛋白完全不溶解，以包涵体的形式出现；而在３０℃诱导表达时，有少部分融合蛋白是可溶的。利用ｇｌｕ－ｔａｔｈｉｏｎｅ－Ｓｅｐｈａｒｏｓｅ４Ｂ对可溶性的融合蛋白纯化后，纯化产物具有抑制黄曲霉孢子萌发的生物活性。Tachyplesin gene was designed and chemically synthesized with the stop codon TAG. The synthesized gene was firstly cloned into vector pUC18 and the recombinant plasmid pFZT2 confirmed by sequencing was obtained. The tachyplesin gene was cloned downstream of GST of prokaryotic expression vector PGEX 5X 1 and the fusion protein GST tachyplesin was induced with 1mmol/l of IPTG. 29 KDa of GST tachyplesin fusion protein band appeared on SDS PAGE. When induced at 37℃, the GST tachyplesin fusion protein was wholly insoluble in aqueous solution; but at 30℃, the fusion protein was partially soluble. After affinity purification of the fusion protein with glutathione Sepharose 4B, the pure GST tachyplesin was used in the tests of fungal inhibition of Aspergillus flavus. The results show that GST tachyplesin fusion protein could inhibit the germination of spores of Aspergilus flavus, but GST could not.

关 键 词：鲎素基因 表达 黄曲霉 大肠杆菌 抑制 

分 类 号：Q78[生物学—分子生物学] TQ455.49[化学工程—农药化工]