摘要
微小毛霉(Mucor pusillus)凝乳酶是微生物凝乳酶的主要来源之一,但与传统的牛凝乳酶比较具有一定的缺陷。为将其采用基因工程的方法进行改造获得理想的凝乳酶,本研究克隆到微小毛霉凝乳酶基因,将其插入原核表达载体pTWlN1中,使之与几丁质结合域(CBD)一内含肽(intein)融合,获得原核表达质粒pTWIN1/M。转化大肠杆菌BL21(DE3),后经IPTG诱导后进行SDS-PAGE电泳分析,获得了重组蛋白。
The rennet from Mucor pusillus is one of the major sources of microbial rennet, but it hassome defects comparing with the bovine chymosin. In order to develop a rennet with suitable milk clotting properties, the gene of rennetwas cloned from Mncor pusilius, and the DNA fragment was inserted into the prokaryotic expression vector pTWIN1, forming pTWIN1/M with a fusion protein integrated with （CBD-intein）. pTWIN1/M was further transformed into E. coli BL21 （DE3）. Expression of the enzyme was performed with IPTG induction. SDS- PAGE tests showed that the rennet from Mucor pusillus was expressed in E. coli BL21 （DE3）.
出处
《中国乳品工业》
CAS
北大核心
2010年第2期7-9,共3页
China Dairy Industry
基金
吉林省科学技术厅资助项目(20060219)
国家863计划探索导向项目(2006AA10Z306)
关键词
微小毛霉
克隆
表达
mucor pusillus
cloning
expression
