hnRPULl和PARP-1的载体构建  

Construction of vectors of hnRPUL1 and PARP - 1.

作　　者：林博[1] 洪泽辉[1] 

机构地区：[1]东南大学医学院,江苏南京210000

出　　处：《中外医学研究》2010年第19期1-3,共3页CHINESE AND FOREIGN MEDICAL RESEARCH

摘　　要：目的构建hnRPULl和PARP-1的原核和真核表达载体，用以表达这些蛋白和研究其相互作用。方法用PCR法扩增hnRPUL1的四个片段基因和PARP-1基因，将扩增产物分别和载体pGEX4T3和pcDNA5进行双酶切后回收，连接载体和目的片段，获得pGEX4T3-hnRPULl和pcDNA5-hnRPUL1的前、中、后、中后四段重组质粒和pGEX4T3-PARP-1重组质粒。然后转化入大肠杆菌DH5ct并进行酶切鉴定和测序。结果hnRPULl和PARP-1基因以正确的阅读框架插入pGEX4T3和pcDNA5。结论成功构建了上述载体并应用于原核和真核表达，为进一步研究其相互作用提供了必要的基础。Objective To construct the vectors of hnRPUL1 and PARP - 1 to for prokaryotic and eukaryotic expression and further stud- ying their interaction. Methods The cDNAs of hnRPUL1 and PARP - 1 were amplified by polymerase chain reaction（PCR） and inserted into pGEX4T3 and pcDNA5 with Xhol and Not1 restriction sites to construct pGEX4T3 - hnRPUL1, pcDNA5 - hnRPUL1 and pGEX4T3 - PARP - 1 respectively. Then the expression plasmids were transformed into E. coli DHSα and positive clones were sequenced. Results The hnRPUL1 and PARP - 1 were inserted into pGEX4T3 and pcDNA5 with correct open read frame. Conclusion The vectors of hnRPUL1 and PARP - 1 were constructed successfully and used both in prokaryotic and eukaryotic expression, which will help us to further study the interaction of hnR- PULl and PARP - 1.

关 键 词：hnRPUL1 PARP-1 载体 构建 

分 类 号：R994.6[医药卫生—毒理学]