文献类型：期刊文献

中文题名：鲑鱼降钙素前体多肽在毕赤酵母中的分泌表达

英文题名：Secreted Expression of Recombinant Salmon Calcitonin Precursor Peptide in Pichia pastoris

作者：季青[1];周祥山[1];张元兴[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2007

卷号：38

期号：6

起止页码：411

中文期刊名：中国医药工业杂志

外文期刊名：Chinese Journal of Pharmaceuticals

收录：CSTPCD;;北大核心:【北大核心2004】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：鲑鱼降钙素;前体;毕赤酵母;分泌表达

外文关键词：salmon calcitonin; precursor; Pichia pastoris; secreted expression

摘要：将鲑鱼降钙素前体多肽(sCTGly)基因克隆到表达载体pPIC9K上,然后转化毕赤酵母营养缺陷型菌株GS115,利用酿酒酵母分泌信号肽α-factor将目的产物分泌到发酵上清液中。经组氨酸缺陷筛选、G418高拷贝筛选及摇瓶表达筛选,获得较高水平表达菌株GS115-sCTGly。酶联免疫反应检测上清液中酶的活性,表明sCTGly产量在10mg/L以上。通过5L发酵罐发酵条件优化,表达水平提高到200mg/L以上。
A gene encoding salmon calcitonin precursor peptide （sCTGly） was cloned into vector pPIC9K and transformated into the methylotrophic Pichiapastoris GS115. sCTGly was expressed and secreted into the culture using the Saccharomyces cerevisiae a-mating factor as signal peptide. Through a series of single-clone screening for His^＋ phenotype, multiple-copy gene insertion and high expression level in shake flask, a strain named GS115-sCTGly with relatively satisfying target characters was obtained. The expression levels of sCTGly were calculated by detecting the activity of the peptide in cell-free supernatants with ELISA, and the yield of sCTGly was over 10mg/L in shake flask culture and could be increased to over 200mg/L by optimization of the fermentation conditions in 5L fermentor.