人成熟型TGF-β_1基因克隆、表达纯化及抗原性的检测  

THE CLONE OF HUMAN MATURE FORM TGF β_1 AND ITS EXPRESSION AND DETECTION OF ITS ANTIGENCITY

作　　者：龚环宇[1] 胡国龄[1] 谭德明[1] 汪玲[1] 侯珏[1] 刘映霞[1] 

机构地区：[1]湘雅医院传染病研究所,长沙410008

出　　处：《中国现代医学杂志》2003年第14期37-40,共4页China Journal of Modern Medicine

摘　　要：目的 :构建人成熟型TGF - β1基因原核表达载体 ,表达并纯化人成熟型TGF - β1融合蛋白。方法 :应用基因重组的方法 ,构建成熟型TGF - β1原核表达载体 ,诱导表达人成熟型TGF - β1的融合蛋白 ,并应用亲和层析法纯化重组融合蛋白 ,应用Western -Blot鉴定其抗原性。结果 :成功构建人成熟型TGF - β1原核表达载体 ,获得人成熟型TGF - β1融合蛋白 ,其抗原性较好。结论 :人成熟型TGF - β1基因原核表达载体的成功构建 ,可高效表达人成熟型TGF - β1融合蛋白 ,为进一步研究其功能和获得其多克隆抗体奠定了基础 ,并为开发国产人TGF - β1诊断试剂做必要的准备 ,对肝纤维化的防治具有较大意义。Objective:The aims of our experiment are to construct the mature form TGF-β 1-PMAL-C2X expression vector, to express the MBP-TGF-β 1 fusion protein, to purify the fusion protein,and to detect its antigencity. Methods:With the recombinant DNA technique,we constructed the mature form TGF-β 1-PMAL-C2X expression vector that was identified by DNA sequence analysis and with restriction enzyme digestion. After induced by IPTG,the MBP-TGF-β 1 fusion protein was expressed in E.coli. The fusion protein was purified by affinity chromatography using amylose resin and testified its antigencity by western-blot.Results:We successfully constructed the mature form TGF-β 1-PMA L-C2X eukaryotic expression vector ,and attained the mature form MBP-TGF-β 1 fusion protein. The mature form MBP-TGF-β 1 fusion protein has good antigencity which was identified by Westem-blot.Conclusions:To construct the mature form TGF-β 1-PMAL-C2X expression vector makes it easy to get the MBP-TGF-β 1 fusion protein. It also can be benifical to study its function and to get the anti-human TGF-β 1 polyclonal antibody. Furthermore,we will try to develop the diagnosis Kit to detect the level of human serum TGF-β 1.

关 键 词：转化生长因子βl 基因表达 基因重组 融合蛋白 

分 类 号：Q755[生物学—分子生物学]