应用RACE法分离和克隆猪GPX2基因研究

摘要

[Objective]Using molecular biotechnology to clone the sus scrofa GPX2 gene.[Method]Using total RNA of sus scrofa duodenum as template,degenerated primer pairs were designed according to the homology alignment analysis of GPX2 gone of human,rat,mouse,dog and cattle.A sus scrofa GPX2 gone sequence of 330 bp was obtained by RT-PCR application method.Primes were designed respectively according to the known sequence,sus scrofa GPX2 gene was isolated and cloned by 3-RACE and 5-RACE method and analyzed the gene sequence.[Result]A mRNA sequence of 924 bp was successfully cloned and isolated in this research.This sequence contained complete 3' end and had higher sequence homology with human,monse,catth and dog GPX2 gene,and there was codon called TGA which encoding See on the position of No.114-116 gene.[Conclusion]Sequence alignment analysis showed that the cloned gene was sus scrofa GPX2 gene(NCBI GenBank database,the sequence number was DQ98982).