摘要
锌指蛋白基因家族是人类最大的基因家族之一,目前已知的很多锌指蛋白成员都是转录调控因子.KRAB/C2H2型锌指蛋白是一类重要的转录因子,ZNF424是该亚家族的一个新成员.已经初步证明ZNF424具有转录激活作用,为了进一步研究ZNF424各结构域激活程度和在信号途径中的作用,设计出引物,以pCMVB-D-ZNF424重组质粒作为模板,PCR扩增出含ZNF424基因的KRAB、LINK和ZNF的3个含不同结构域的区域,克隆到pMD18T-载体,然后内切酶切下各目的片段,再构建出pCMVB-DK-RAB、pCMVB-DL-INK、pCMVB-D-ZNF、pCMVT-ag2BK-RAB、pCMVT-ag2BL-INK和pCMVT-ag2CZ-NF共6个缺失突变重组质粒,为进一步研究ZNF424基因功能奠定基础.
Zinc finger protein family is one of the largest families and lots of the members are the transcriptional regulators.The type of KRAB/C2H2 zinc finger protein is a significant kind of transcriptional regulate factors,and a novel human ZNF424 belongs to this sub-family.In order to examine the function of each domain of ZNF424 in the transcriptional activity and in the signal pathway.Different primers for KRAB,LINK and ZNF were designed.PCR was performed using the pCMV-BD-ZNF424 as the template according to standard procedures.Then each PCR product was ligated into pMD18-T vector.Finally,pCMV-BD-KRAB,pCMV-BD-LINK,pCMV-BD-ZNF,pCMV-Tag2B-KRAB,pCMV-Tag2B-LINK and pCMV-Tag2C-ZNF deletion mutation recombinant plasmids were constructed,which established the foundation for further examing the function of ZNF424 gene.
出处
《生命科学研究》
CAS
CSCD
2006年第S2期1-9,共9页
Life Science Research
基金
国家自然科学基金资助项目(30470867
90508004
30270722
30570934
30571048
30570265)
湖南省教育厅项目资助项目(04C327)
关键词
锌指蛋白
ZNF424
缺失突变
重组质粒
zinc finger protein
ZNF424
deletion mutation
recombinant plasmid
