摘要
①目的 利用基因工程技术从人扁桃体细胞cDNA中克隆出白细胞介素 2 1(IL 2 1)的编码基因 ,并在大肠杆菌中进行表达。②方法 以人扁桃体细胞经PHA刺激的cDNA文库为模板 ,经PCR获得IL 2 1的全基因片段。测序确认后 ,分别设计含BamHⅠ和SalⅠ酶切位点的引物 ,经PCR获得IL 2 1成熟肽的编码基因。将此IL 2 1基因插入表达载体 pGEX4T 2 ,重组克隆经酶切与测序确认后转化大肠杆菌DH5α进行IPTG诱导表达。 ③结果 琼脂糖凝胶电泳显示重组质粒的PCR和双酶切产物与理论值的大小相等。SDS PAGE显示经IPTG诱导后的大肠杆菌DH5α亦表达一与理论相符的条带。目的蛋白约占总菌体蛋白的 10 %。④结论 成功地构建了IL 2
Objective Using gene engineering technique, IL 21 code gene was cloned from cDNA of human tonsil cells and expressed in E.coli.\ Methods\ Human interleukin 21 gene fragment was acquired from human tonsil cDNA by means of PCR. The matured N terminus of gene code of IL 21 was inserted into prokaryotic expressing vector pGEX4T 2 and transformed into E.coli DH5α. \ Results\ The electrophoresis showed plasmid PCR and corresponding products of BamH Ⅰ and Sal Ⅰ were the same as expectation. After inducing by IPTG, E.coli DH 5α also expressed the right molecular weight protein. The objective protein accounted for about 10% of the whole bacterial protein.\ Conclusion\ The vector of pGEX4T 2/IL 21 is constructed and expressed successfully in E.coli.
出处
《青岛大学医学院学报》
CAS
2003年第1期15-17,共3页
Acta Academiae Medicinae Qingdao Universitatis
基金
山东省自然科学基金重点资助项目 (E99C0 2 )
