原核表达基质金属蛋白酶-2的催化域

Prokaryotic expression of catalytic domain of matrix metalloproteinase-2

目的 体外表达人基质金属蛋白酶 - 2 (MMP - 2 )的催化域 ,为进一步研究其在肿瘤侵袭和转移中的作用打下基础。方法 通过PCR方法获得了编码MMP - 2催化域 (MMP - 2 -C)的cDNA序列 ,并构建了重组质粒pGEX - 5X - 3/MMP - 2-C ,由IPTG诱导在大肠杆菌中表达 ,表达产物被复性后鉴定酶活性。结果 表达产物为 6 8-Kda的融合蛋白且具有明胶酶活性。

Objective Expression of the catalytic domain of human matrix metalloproteinase 2(MMP 2) will faciliate the study on tumor invasion and metastasis.Methods The cDNA sequence of encoding the catalytic domain of human MMP 2 was obtained by polymerase chain reaction (PCR).The prokaryotic expression vector containing the catalytic domain of matrix metalloproteinase 2(pGEX 5 X 3/MMP 2 C) was constructed.Expression products were obtained from transfected E.coli induced by IPTG.The fusion protein was renatured,and its activity was detected.Results The expression product 68 Kda fusion protein has the activity of gelatinase.Conclusions The in vitro expressed fusion protein with catalytic activity can be used for studying drugs against tumor invasion and metastasis.