人基质金属蛋白酶-1的基因克隆、表达及其产物抗原性的分析  

Cloning and expression of the human MMP-1 gene fragment and antigenecity analysis of its expression products

作　　者：侯珏[1] 胡国龄[1] 刘双虎[1] 谭德明[1] 欧阳颗[1] 

机构地区：[1]中南大学湘雅医院传染病研究所,长沙410008

出　　处：《中华传染病杂志》2003年第4期249-252,共4页Chinese Journal of Infectious Diseases

摘　　要：目的　构建人基质金属蛋白酶 1(MMP 1)基因重组质粒的克隆 ,获得具有抗原性的人MMP 1融合蛋白。方法　从人肝组织提取总RNA ,以此为模板 ,逆转录巢式PCR扩增MMP 1全编码区基因片段 ,构建含目的片段的T载体克隆及原核表达载体 pMAL c2x重组质粒亚克隆 ,经异丙基 β D半乳糖苷酶 (IPTG)诱导表达MMP 1重组质粒菌 ,用十二烷基硫酸钠聚丙烯酰胺凝胶电泳 (SDS PAGE)和Western blot对重组蛋白进行分析鉴定。结果　经核苷酸序列测定和免疫印迹鉴定 ,成功地构建了人MMP 1重组质粒基因工程菌 ,并表达具有MMP 1抗原性的融合蛋白。结论　构建了人MMP 1重组质粒克隆 ,获得了具有抗原性的MMP 1融合蛋白 ,这将有助于今后制备MMP 1多克隆抗体。Objective To construct the recombinant plasmid of human MMP 1 clone and study its antigenecity of MMP 1 fusion protein. Methods The total RNA was extracted from human liver and used as a template for reverse transcription. After PCR amplification, a 1 432 bp fragment was obtained and cloned into T vector. After digested with restriction enzyme, the target fragment was subcloned into plasmid pMAL c2x. The recombinant plasmid was transferred into JM109 which can express a fusion protein. We analyze the protein with SDS PAGE and Western blot. Results We obtained human MMP 1 gene and its recombinant plasmid clone. The expressed protein can be recognized by MMP 1 polyclonal antibody in Western blot.Conclusions We obtained the MMP 1 protein with antigenicity. The fusion protein can be used to prepare polyclonal antibody against MMP 1.

关 键 词：人基质金属蛋白酶-1 基因克隆 基因表达 原核表达 抗原性 T载体克隆 肝纤维化 诊断 

分 类 号：R346[医药卫生—基础医学]