人核心蛋白聚糖真核表达载体的构建及鉴定

目的：构建人核心蛋白聚糖（DCN）真核表达载体，为进一步研究其生物学活性奠定基础。方法：采用聚合酶链反应（PCR）扩增目的片段，PCR产物及真核表达载体pcDNA3分别双酶切后进行连接，并转化入大肠杆菌JM109中扩增以获得重组载体。结果：PCR获得与预期大小一致的、约1 000 bp的特异性DNA片段，PCR产物与表达载体经双酶切后连接，构建重组载体pcDNA-dec，经双酶切鉴定及测序证实，人DCN基因cDNA片段正确插入真核表达载体中。结论：成功构建人DCN真核表达载体pcDNA-dec。

Construction and identification of human decorin gene recombinant eukaryotic expressing vector

Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin(DCN).Methods DCN cDNA was amplified by using polymerase chain reaction(PCR).Product of PCR and expressing vector were digested by restriction endonucleases,then ligated and transformed into JM109 bacteria.Results The specific DNA fragment was obtained by PCR as supposed.Product of PCR and expressing vector were digested by restriction endonucleases,then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec.It was confirmed that DCN cDNA was inserted into the eukaryotic expression vector correctly by using digestion identification and sequencing.Conclusion The recombinant eukaryotic expression vector pcDNAdec of human DCN is successfully constructed.