Genetic Evolution Analysis of S1 Gene of a PEDV Guangdong Epidemic Strain and Its Expression and Purification in Insect Cells
In order to understand the prevalence and variation of porcine epidemic diarrhea virusS1 gene of Guangdong epidemic strains corresponding diagnostic reagents were prepared.The S1 gene was amplified by RT-PCR from intestinal contents of piglets with suspected epidemic diarrhea,and the S1 genetic evolution tree was constructed by bioinformatics software.Then S1 gene was cloned into baculovirus shuttle plasmid pFastBac1,and transformed into DH10Bac competent cells.After screened by antibiotics,the recombinant baculovirus plas-mid was transferred into insect cell Sf9.After 3 generations of blind transmission,IFA test and western blots iden-tification were carried out.The results show that GD2020 strain was located in GII-b branch,and the nucleotide similarity with domestic PEDV isolates(HN-HB1-2018,HB-HA2015)is 98.0%～98.3%,but the nucleotide simi-larity with the classical strains CV777 and DR13 is only 89.9%～90.0%.And S1 recombinant protein can be suc-cessfully expressed in insect cell Sf9 in the form of supernatant secreted by cell.This research can provide a refer-ence for the epidemic status of PEDV,and provide early foundation for the research and development of diagnos-tic products of porcine epidemic diarrhea virus.
万方数据
维普
