毕赤酵母碳源阻遏因子编码基因PpMIG1和PpMIG2的克隆及序列分析.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 11 PpM IG1 PpM IG2 张平 周祥山 张元兴 ( , 200237) : 通过M IG基因的同源性设计简并引物, 采用 PCR 方法从毕赤酵母(P ich ia p astoris ) 中 功克隆了两 个M IG 同 源基因的核心片段, 同时利用 Genom eW alk ing的方法获得了基因的全长及其侧翼序列, 并分别命名为 PpM IG 1和 PpM IG2序 列分析显示, PpM IG 1基因全长 1 335 bp, 编码 444 个氨基酸; PpM IG2基因全长 1 365 bp, 编码 454 个氨基酸这 个两 个基因所 编码的蛋白质均与酿酒酵母碳源阻遏因子 cM ig1同源, 在氨基末端具有两 个典型的 C H 锌指结构域, 该结构域能够与受葡 2 2 萄糖阻遏的许多基因的启动子相结合PpM ig阻遏因子的获得为毕赤酵母碳源阻遏机制的研究奠定了基础 : 毕赤酵母 碳源阻遏因子 克隆 Cloning and Sequence Analysis of the Genes PpM IG1 and PpM IG2 Encoding Catabolite Repressors inP ichia p astoris Zhang Ping Zhou X iangshan Zhang Yuanx ing (S ta teK ey La boratory of B ioreac tor E ng ine ering, E ast China University of S cience and T echnology, Shangha i 200237) Abstrac:t In th is study, w e have cloned two M IG1 hom ologous genes inP ich ia p atoris based on degenerate PCR and Genom e W alk ing m ethod. The tw o genes have been designated PpM IG 1 and PpM IG2, respectively. equence analysis of the genes revealed a 1 335 bp open read ing fram e coding for 344 am ino acids in the case of PpM IG 1, and a 1 365 bp open reading frame for 354 am ino acids in the case of PpM IG2. Both Pp M IG genes encode protein hom ologous to the DNAb inding repressorM ig1 from S accharom yces cerev isiae ( cM ig1), wh ich is a C H zinc finger protein and could bind to the promoters of m any genes repressed by glucose. The 2 2 find ing of these two catabolite repressors is the first step in elucidating them echanism s of catabolite repression inP p astoris. K ey w ords: P ich ia p astoris