基质金属蛋白酶-2C端片段pex的克隆、测序及真核表达载体的构建

Cloning and Sequencing of Matrix Metalloproteinase-2 C End Segment Pex and Construction of its Eucaryotic Expression Vector

目的 :克隆小鼠基质金属蛋白酶 - 2 ( MMP- 2 ) C端片断 pex编码区的 c DNA序列 ,并构建其真核表达载体 ,为进一步研究其抑制肿瘤作用奠定基础。方法 :根据基因 Gen Bank中 MMP- 2基因的碱基序列 ,利用 RT- PCR的方法从 NIH3T3细胞中分别扩增信号肽及目的基因 pex,然后用多重 PCR的方法将两个片段拼接起来 ,再将拼接起来的目的片段 sig- pex与 p MD1 8T载体连接作全自动测序 ,利用亚克隆的方法将 sig- pex c DNA片段克隆到 pc DNA3.1载体中。结果 :DNA测序证实该片断序列与文献报道完全一致。结论 :成功构建出 pc DNA3.1 - sig-

Objective:To clone the sequence of cDNA of matrix metalloproteinase of mouse the sequence and construct its expression vector, provide a novel approach for the study of tumor growth inhibition.Methods:In the GenBank,the signal peptide gene and the aim gene pex from the NIH3T3 cells were amplified using RT PCR,respectively. The two gene segments were combined by means of multiple PCR,Finally, the combined aim gene segment Sig pex connected to pMD18T vector was subjected for auto sequencing. Results:DNA sequence conformed that the sequence of the analyzed gene segment was in consistent with the one formerly reported in the literature.Conclusion:The eukaryotic expression vector that derived from Sin pex cloned into pcDNA3.1 vector has been completed successfully. [