摘要
目的 获得足量的PDGF BB蛋白 ,作为PDGF BB促进骨折愈合及创伤修复等进一步功能研究和临床应用的基础。方法 用基因重组技术构建pQE PDGF B原核表达载体 ,在M15大肠杆菌中进行PDGF B单体蛋白的表达。结果 经酶切鉴定、PCR鉴定及基因测序证明 ,pQE30载体上成功地插入了PDGF B成熟肽基因 ;重组表达载体pQE PDGF B在M15大肠杆菌中得到了高效表达 ,表达量约占全菌蛋白的 15 % ,表达的PDGF B单体蛋白经SDSPAGE电泳 ,显示了一条特异蛋白带 ,分子量为 15kD左右。结论 PDGF B原核表达载体的成功构建和PDGF B单体蛋白制备纯化为生产活性PDGF BB蛋白及其进一步功能研究奠定了基础。
Objective To acquire sufficient PDGFBB protein and provide the basis for the further studies of its role on the fracture healing and trauma reconstruction and its clinical applications. Methods Constructed the prokaryotic expression vector pQEPDGFB with the gene rearrangement technique, and the monomeric form of recombinant PDGFB expressed in E. coli M15. Results PDGFB mature peptide gene was inserted into prokaryotic expression vector pQE30,which was confirmed by PCR、enzyme digestion and sequencing identification;the expressed products of pQEPDGFB in E. coli showed a single protein on SDSPAGE,and their expression level was about 15% of the total bacterial protein。The molecular weight of the purified PDGFB protein was about 15 KDs on SDSPAGE. Conclusions The construction of recombinant plasmid and preparation of the monomeric protein of PDGFB provides a solid foundation for further studying the function of PDGFBB and producing biologically PDGFBB protein.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2004年第19期1170-1173,共4页
Chinese Journal of Surgery
基金
国家自然科学基金资助项目 (3 0 2 713 2 1)
