人源性基质金属蛋白酶-1原核表达载体的构建与鉴定

Construction and Identification of the Prokaryotic Expression Plasmid of Human MMP-1 Gene Fragment

目的 构建人源性基质金属蛋白酶 -1(MMP -1)原核表达载体。 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 ,经双酶切及核苷酸测序进行分析鉴定。 结果 成功地构建了人MMP -1原核表达载体。 结论 构建的人MMP -1原核表达载体 ,为以后MMP -1融合蛋白的表达并获得具有抗原性的MMP -1融合蛋白奠定了基础。

Objective To construct the prokaryotic expression vector of human MMP-1 gene fragment. Methods The total RNA was extracted from human liver tissue and used as a template for reverse-transcription. After PCR amplification,a 1432bp fragment was obtained and cloned into T vector. After digested with restricted enzyme,the target fragment was subcloned into plasmid pMAL-c2x,the recombinant plasmid was transferred into JM109. We analyze the fragment with restricted enzyme and nucleotide sequencing. Results We obtained human MMP-1 gene and its prokaryotic expression vector. Conclusion We obtained the prokaryotic expression vector of human MMP-1 gene fragment ,which will further help in making MMP-1 fusion protein and the polyclonal antibody against MMP-1.