表达人胰岛素前体的毕赤酵母菌株的构建

Construction of recombinant HIP Pichia pastoris strains

将人胰岛素前体 (HIP)基因插入到毕赤酵母Pichiapastoris的分泌表达质粒 pPIC9K中 ,得到分泌表达质粒pPIC9K/HIP并用电转化法转化P .pastorisGS1 1 5。筛选出整合型His+ Muts 菌株 ,进一步用G4 1 8筛选获得高拷贝转化子。经诱导培养后 ,SDS 聚丙烯酰胺凝胶电泳 (SDS PAGE)证明HIP在毕赤酵母中能有效分泌表达。对毕赤酵母中外源基因的稳定性进行了研究 。

The human insulin precursor (HIP) gene was inserted into the plasmid pPIC9K of Pichia pastoris to obtain secretory plasmid pPIC9K/HIP.After electroporation of Pichia pastorisGS115(His -Mut +),some high copy transformants (His +Mut s) were picked up by G418.SDS PAGE analysis indicated the efficient expression and secretion of HIP. The stability of HIP gene in the recombinant Pichia pastoris was studied and the results showed that the recombinant plasmid had good stability in Pichia pastoris.