人肥大细胞类糜蛋白酶分泌型真核表达载体的构建

目的构建人肥大细胞类糜蛋白酶（MC—Chy）分泌型真核表达载体。方法以MC-Chy基因的质粒pDEST17／CMA为模板，通过PCR扩增出融合有免疫球蛋白K链信号肽的人MC—Chy基因，定向克隆至真核表达载体pcDNA3．1，通过PCR、酶切和DNA测序鉴定。结果PCR扩增出免疫球蛋白K链融合人MC-Chy基因，DNA序列测定结果显示目的基因片段正确插入真核表达载体pcDNA3．1。结论成功构建了人MC-Chy分泌型真核表达载体，为下一步重组类糜蛋白酶的制备及其功能的深入研究奠定了基础。

Construction of the secretory type recombinant human mast cell chymase gene eukaryotic expression vector

Objective To construct the secretory type recombinant human mast chymase （MC-Chy）gene eukaryotic expression vector. Methods DNA fragment encoding human MC-Chy fusion to the signal peptide of immunoglobulin kappa-light-chain was amplified by PCR, then was subcloned the eakaryotie expression vector pcDNA3.1, which was identified hy PCR, digestion of enzyme and DNA sequencing. Results DNA fragment encoding human MC-Chy fusion to the signal peptide of immunoglobulin kappa-light-chain was amplified by PCR, and which was inserted into the eukaryotle vector pcD- NA3.1. Conclusion The secretory type recombinant human MC-Chy gene eukaryotic expression vector was constructed successfully, which lays the foundation for further preparing the recombinant human MC-Chy by eukaryotic expression and its function research.