摘要
周期素依赖性激酶抑制蛋白（ｃｙｃｌｉｎ－ｄｅｐｅｎｄｅｎｔｋｉｎａｓｅｉｎｈｉｂｉｔｏｒｐｒｏ－ｔｅｉｎ，ＣＫＩ），如ｐ２７和ｐ１５基因是抑癌基因的候选基因。作者通过ＰＣＲ方法获得含人ｐ２７全部编码区域的ｐ２７ｃＤＮＡ片段，并用ＰＣＲ产物ＴＡ克隆法，将其构建在真核表达载体ｐＣＲＴＭ３中；用常规酶学方法，通过ＥｃｏＲＩ和ＸｈｏＩ双酶切位点，将人ｐ１５ｃＤＮＡ片段克隆入ｐＣＲＴＭ３载体。重组体的鉴定通过限制酶图谱分析进行。人ｐ２７和ｐ１５基因真核表达重组体的构建为进一步研究其功能打下基础。
yclindependent kinase inhibitors(CKIs),such as p27 and p15,are potential candidates for tumor suppressors. A cDNA fragment containing the complete coding region for human p27 was prepared by polymerase chain reaction and ligated efficiently with the linearized expression vector pCRTM3 through a new cloning technique of PCR productsEukaryotic TA Cloning System. The EcoR I/Xho I site of human p15 cDNA was inserted into the EcoR I/Xho I site of pCRTM3 vector through conventional enzymatic method. Two recombinants were identified by restriction analysis. By the construction of eukaryotic expression vectors for human p27 and p15 we made the functional study of this two genes possible.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
1997年第5期441-444,共4页
Journal of China Medical University
基金
中华医学基金
