鲑鱼降钙素基因在大肠杆菌中的可溶性高效表达  

Soluble and High Expression of Salmon Calcitonin Gene in E.coli

作　　者：刘深基[1] 陈松森[1] 张松[1] 狄旭[1] 

机构地区：[1]中国医学科学院

出　　处：《基础医学与临床》1999年第3期89-93,共5页Basic and Clinical Medicine

摘　　要：用PCR的方法从含有鲑鱼降钙素基因的质粒中扩增出目的基因，并将其插入硫氧还原蛋白融合表达质粒pTrxFus中，转入大肠杆菌GI724，以色氨酸诱导，使之与硫氧还原蛋白融合表达，融合蛋白可占菌体总蛋白的50％左右。用超声破碎菌体后，上清直接上CM－SepharoseFF柱进行分离纯化，得到融合蛋白的纯度达90％以上。动物体内活力分析表明，融合蛋白有明显的降低血钙的作用，此工作为重组鲑鱼降钙素的开发应用和功能研究打下了基础。The sCT gene was obtained from plasmid pGEX-sCT by PCR amplification. After treated with Klenow and Xba I, It was inserted into plasmid pTrxFus through polyclonal sites Sma I and Xba I to construct the expressing plasmid pTrx-sCT. The expressing plasmid was transferred into E.coli K12 strand GI724 and induced with tryptophan to express the fusion protein of thioredoxin and salmon calcitonin, SDS-PAGE results show that the recombinant fusion protein can reach 50 % of the total bacteria protein and are mostly in soluble form. Disrupted by sonication and then centrifuged,the supernatant was separated by CM-Sepharose FF ion exchange chromatography eluted with linear gradient NaCl. The fusion protein whose purity was above 90 %. The results of experiments in vivo show that the recombinant fusion protein can lower the level of serum calcium significantly.

关 键 词：鲑鱼降钙素 降钙素基因 可溶性 高效表达 

分 类 号：Q78[生物学—分子生物学] Q572