摘要
目的 :为获得高表达有活性的p16蛋白 ,构建含p16cDNA的重组转移载体。方法 :EcoRI xhoI双酶切克隆质粒pBLUESCRIPT p16 ,低融点琼脂糖回收 0 8kb的p16cDNA片段 ,插入质粒pSXIVVI+X3,构建含p16的重组载体pX3 p16 ,并对重组子以菌落原位杂交和酶切电泳两种方法进行鉴定。结果 :电泳证实低融点琼脂糖回收的片段为 0 8kb。菌落原位杂交筛选 8个阳性克隆 ,酶切电泳证实阳性克隆中p16cDNA插入方向正确。结论 :成功构建了含p16cDNA的重组转移载体 。
Objective:To get plenty of active p16 protein,and build a recombined transfer vector with p16 cDNA. Methods:p16 cDNA carried by cloning vector pBLUESCRIPT p16 was cut down with EcoRI and xhoI,then collected and purified by low melting agarose.The 800bp long p16 cDNA was inserted into transfer vector carrying p16 cDNA(pX 3 p16)was selected and identified by in situ hybridization and electrophoresis. Results: 800bp of p16 cDNA purfied from low melting agarose war assayed by electrophoresis.8 positive clones were obtanined after in situ hybridization with p16 cDNA probe,and the transfer vector carrying with p16 cDNA(pX 3 p16)was identified by digestion of EcoR I and xhoI. Conclusion:pX 3 p16 was a recombiant of p16 cDNA and pSXIVVI +X 3.
出处
《山东医科大学学报》
2000年第3期243-244,共2页
Acta Academiae Medicinae Shandong
基金
卫生部科研基金资助课题! (96 2 1 5 8)
关键词
抑癌基因
P16基因
转移载体
构建
Genes,suppressor,tumor
Genes,p16
Transfer vector
Baculoviridac
