人β干扰素血清白蛋白融合蛋白在毕赤酵母中的分泌表达.pdf

·2006年第六届国际酒文化学术研讨会论文集· 人p干扰素一血清白蛋白融合 蛋白在毕赤酵母中的分泌表达 雷楗勇， 张莲芬， 杨健良， 金 坚‘ (江南大学生物工程学院教育部工业微生物重点实验室， 无锡214036) 导袁迭，SDS—PAGE和Western 子量约为90kDa且具有HsA的抗原性；用细胞病变抑制法测定发酵液上清中融合蛋 白的干扰素活性约为6401U／mL。 关键词：口干扰素；巴斯德毕赤酵母f融合蛋白 oftheFusionProtein SecretoryExpression inPichia IFNp’HSA pastoris LEI ZHANG YANG JINJian Jian-Yong， Lian-Feng， Jian-Liang， (The ofIndustrial ofEducation．Schoolof KeyLaboratory Biotechnology，Ministry Southern China 214036) YangtzeUniversity，Wuxi．PR PCR was to HSA Abstract：OverlappingtechnologyemployedspliceIFN口andgenes invitro．The wasinsertedintoPichia vector splicedgene pastorissecretory was the tobe under pPIC9K．The gene designedsecretoryexpression IFNl3一HSA controlof AOXlandMatd in promoter signalpeptidepPIC9K．The plasmid was1inearizedrestriction SalIand pPIC9K／IFN[3一HsA by enzyme transformedintoPichia KM71 recombinantstrains pastorisbyeleetroporation．The identified and G418selectionconformedPCR wereinduced by by analysis by methanolto fusion andWesternblot express protein IFNfl—HSA．SDS—PAGE ofthefusion showedthat