丝状真菌表达分泌系统中受体菌的构建  

Construction of Recipient Strain of Expression-secretion System in Filamentous Fungi

作　　者：刘丽[1] 刘谨[2] 仇润祥[1] 朱兴国[1] 唐国敏[1] 

机构地区：[1]中国科学院微生物研究所,北京100080 [2]山东大学,济南250001

出　　处：《生物工程学报》2002年第6期667-670,共4页Chinese Journal of Biotechnology

基　　金：国家自然科学基金资助项目 (No.30 170 0 14 )~~

摘　　要：黑曲霉糖化酶高产菌株T2 1经紫外诱变后 ,通过酪蛋白平板和蛋白酶活性测定筛选出胞外酸性蛋白酶活力仅为原株 0 76 %的菌株A .nigerT2 1 2 0 1,其生长特性和产糖化酶活力与原株基本一致。利用原生质体 PEG法将含有报告基因vhb的表达分泌质粒pGT10 vhb通过与选择标记质粒的共转化导入此蛋白酶部分缺陷株及其原株T2 1,检测在蛋白酶缺陷株AspergillusnigerT2 1 2 0 1和原株T2 1中VHb的分泌表达 ,结果表明在A .nigerT2 1 2 0 1中VHb表达水平显著高于原株 ,但Northernblot却显示在两菌株中vnb基因的转录水平近似 。Glucoamylase overproducing A.niger T21 was mutated by UV mutagensis. An extracellular acid protease-deficient mutant, A.niger T21-201, which produced only 0\^76% extracellular acid protease activity of the parent strain, was screened by casein-degradating plate and determination of protease activity. Morecover, the growth properties and the ability to secrete glucoamylase of A.niger T21-201 are identical to these of starting strain T21. The comparison of expression-secretion levels of heterologous gene in A.niger T21-201 and T21 was carried out with bacterial vhb as reporter. the level of expression-secretion of VHb in A.niger T21-201 was 6～7 times higher than that in T21, but the transcriptional levels of vhb gene in both strains were similar revealed by Northern blot. Therefore, it was demonstrated that the deficiency of acid protease of recipient T21-201 has significant effect on the protection of heterologous protein.

关 键 词：丝状真菌 表达分泌系统 受体菌 构建 黑曲霉T21 蛋白酶缺陷株 VHB 

分 类 号：Q782[生物学—分子生物学]