中国药科大学学报2005，Vol.36Issue(6)：590-593,4.
一种合成人血小板因子4基因的简便有效方法
A Convenient and Effective Approach for Gene Synthesis of Human Platelet Factor-4
摘要
Abstract
AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.关键词
人血小板因子4/基因合成/基因表达利用纳米技术运送抗癌药的新给药方式Key words
Human platelet factor-4/Gene synthesis/Gene expression分类
生物科学引用本文复制引用
吴晓萍,苏志坚,郑青,吴思娴,许华,赵文,李校堃..一种合成人血小板因子4基因的简便有效方法[J].中国药科大学学报,2005,36(6):590-593,4.基金项目
国家高技术研究发展计划("八六三"计划)资助项目(No.2002AA2Z3318) This work was supported by the National High Technology Researeh and Development Program of China( No.2002AA2Z3318) （"八六三"计划）
