民猪ZFAND5基因的克隆与真核表达载体构建

根据NcB1上已公布野猪的ZFAND5序列,设计上下游引物,PCR扩增,构建真核表达载体,酶切鉴定并测序.克隆并分析ZFAND5(zinc finger,AN1-type domain5)基因序列,构建zFAND5基因的真核表达载体,为转染细胞提供基础.通过研究首次获得了民猪的ZFAND5基因的全序列及其完整的cDNA...

Molecular cloning and construction of eukaryotic cell expression vector of ZFAND5 gene in Min pig

Making a study of sequence analysis of ZFAND5(zinc finger,AN1-type domain5) and the construction of the eukaryotic cell expression vector of pcDNA3.1(-)-ZFAND5 to lay the foundation for the cell transfection.The primers were designed according to the sequence of ZFAND5 gene,PCR,constructed eukaryotic cell expression vector,then restricted enzymes analysis and sequenced.This study was the first time to get the Min pig's complete sequence of DNA and cDNA of ZFAND5 gene,by means of sequence alignment,4 point mutations were found in the two exons of ZFAND5 gene,in which the 643 point causes a deletion of HhaII restriction site.Restricted enzymes analysis and DNA sequencing showed that the sequence of the recombinant plasmid pcDNA3.1(-)-ZFAND5 was correct.These results showed the function and polymorphism of ZFAND5 gene and the accomplishment of the construction of eukaryotic cell expression vector of pcDNA3.1(-)-ZFAND5,laying the theoretical foudation for the tesearch expression and function of ZFAND5 gene in cells.