毕赤酵母表达人溶菌酶基因的初步研究  

Preliminary studies on expression of human lysozyme gene in Pichia pastoris

作　　者：王安平[1] 朱善元[1] 王永娟[1] 吴双[1] 左伟勇[1] 洪伟鸣[1] 

机构地区：[1]江苏畜牧兽医职业技术学院,江苏省兽用生物制药高技术研究重点实验室,江苏泰州225300

出　　处：《江苏农业学报》2013年第3期574-577,共4页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省自然科学基金项目(BK2011536);院重点项目(ZD201105)

摘　　要：为在毕赤酵母中表达人溶菌酶基因,根据已发表的人溶菌酶基因序列设计引物,扩增人溶菌酶全基因片段,将其克隆进巴斯德毕赤酵母分泌型表达载体pPIC9K中,构建重组质粒pPIC9K-LYZ,经Sac I酶切线性化后电转化毕赤酵母GS115,经G418筛选阳性克隆,并用甲醇诱导表达。SDS-PAGE证实表达的重组蛋白质分子量正确,抑菌试验结果表明重组hLYZ对微球菌具有良好的抑菌效果。说明人溶菌酶基因在毕赤酵母中成功表达。In order to express human lysozyme （hLYZ） gene in Pichia pastoris, one pair of primers was designed according to the published sequences for hLYZ, and the hLYZ gene was amplified by PCR and cloned into the yeast secretory expression vector pPICgK to construct the recombinant plasmid pPIC9K-LYZ. The recombinant plasmid was transformed to P. pastoris GS115 after digested by restriction enzyme Sac I, and positive clones were screened with G418 resistance for expression induced by 0.5% methanol. SDS-PAGE confirmed the correct molecular weight of recombinant expression protein. Antibacterial assay showed that hLYZ had obvious inhibitory activity against micrococcus. These resuhs indicate that hLYZ gene has been expressed successfully in P. pastoris.

关 键 词：人溶菌酶 毕赤酵母 表达 抑菌活性 

分 类 号：S859.797[农业科学—临床兽医学]