摘要
目的克隆编码人赖氨酰氧化酶(hLOX)的基因片段,并构建含有该基因的重组原核表达质粒,为制备基因工程化的重组hLOX奠定基础。方法从人皮肤成纤维细胞中抽提RNA,通过RT-PCR扩增LOX编码序列(750bp),所得目的基因插入原核表达载体质粒pET28a,转化大肠杆菌DH5α。重组质粒pET28a-hLOX经双酶切和核苷酸序列分析鉴定。结果载体质粒pET28a中插入的基因片段与hLOX基因序列完全相同。结论成功构建含有人LOX基因的重组表达质粒。
Objective To clone gene encoding human Lysyl oxidase（hLOX）and construct prokaryotic expression plasmid which lay the foundation for expressing the recombinant hLOX in the future. Methods Total RNA was extracted from human skin flbroblast cells and the gene fragment of hLOX（750bp） was obtained with RT-PCR. The segment was inserted into prokaryotic vector pET28a and the inserting plasmid was transformed into Escherichia coli. host DH5a. The positive clone was characterized by restriction endonuclease mapping and DNA sequence analysis. Results The gene encoding hLOX was inserted into vector pET28a accurately. Conclusion The recombinant prokaryotie expression plasmid pET28a-hLOX was successfully constructed by properly inserting gene encoding hLOX.
出处
《宁夏医科大学学报》
2009年第4期421-422,425,F0002,共4页
Journal of Ningxia Medical University
基金
宁夏自然科学基金
项目编号:NZ0772
宁夏医科大学校级课题(2005)
