Title: Cloning of Human PCK1 Gene and Construction of Its Expression Vector
Abstract: Objective To construct a eukaryon expression vector of human PCK1 gene. Methods The whole coding sequence of PCK1 gene was amplified by polymerase chain reaction(PCR) applied to human adipose cell(cDNA.) The fragment was inserted into cloning vector pGEM-T and the recombinant pGEM-T-PCK1 was identified by double digestion with restriction enzymes and sequencing.The PCK1 cDNA fragment was then subcloned into(pcDNA3.1(+)) plasmid.The recombinant pcDNA3.1-PCK1 was identified by double digestion with restriction enzymes.(Results)The length of the specific fragment by PCR was 1 972 bp,and the pGEM-T-PCK1 plasmid was(identified) by endonuclease digestion and sequencing.The sequence was identical to that of PCK1 cDNA in GenBank.The recombinant(pcDNA3.1-PCK1) plasmid was separated into two bands,namely 5.1 kb and 2 000 bp,by using respective restriction enzymes Kpn Ⅰ/Xba Ⅰ,suggesting that PCK1 gene fragment had been cloned into pcDNA3.1 vector correctly.(Conclusion)The wild-type recombinant eukaryon expression vector pcDNA3.1-PCK1 was successfully constructed.
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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