Cloning And Bioinformatics Analysis Of GC Gene Of Pseudorabies Virus

We got DNA of Pseudorabies virus Fa strain, vaccine strain 783, Bartha and SA215, Sichuan wild strain SN, SS, SQ, SL and SCZ preservatived by Animal Biotechnology center of Sichuan Agriculture University, and Beijing BJ strain, Guangdong DG strain, Hubei HS strain donated by prof. Lou Gaoming of Shao Guan Institute Ying Dong Bioengineering College. DNA of the 12 Pseudorabies virus strains served as template, 12 strips of gC gene fragments of PRV were amplified by PCR. Reclaimed the product of PCR, ligated and allaxised into competent cell of E.coli DH5α, then sent to biology company to sequencing after accredited by colony PC R and biocatalyst cutting.The 12 strips gC gene sequences of different PRV strains, as well as 6 strips gC genes sequences which were downloaded from GenBank were analyzed by bioinformatics software. Analyzed and predicted that, such as nucleotide homology, location of mutation area, relationship of heredity and evolution, discrepancy of biocatalyst cutting site, preference of codon, homology of amino acid sequence, protein hydrophilicity, epitope, secondary structure and tertiary structure. The results showed that the length of open reading frame of gC gene was between 1437bp and1464bp, the number of amino acid encoded correspondingly was between 478 and 487. The ribonucleotide autoploidy was between 94.9% and 100%, the deduced autoploidy of amino acid sequence was between 90.2 % and 100%.There was a concentrated mutation area and a nucleotide deletion area on the 175-222 location segment of the nucleotide sequence, 8 basic groups mutated and 21 basic groups inset or absented. To analyze on the heredity and evolution relationship, the genetic relationship of the pseudorabies vims strains prevalent in Sichuan were closer to the pseudorabies vims strains isolated in Japan, Europe and America, but distant to the strains isolated in some region of our country. There was no HindIII biocatalyst cutting site in all 18 strips of gC gene, and there was no BamHI biocatalyst cutting site in Fa strain, DG strain ,SA215 strain ,BJ strain ,Min A strain and Ea strain either.Yamagata S-81 strain had no SalI site. There were tiny discrepancy on the occurrence frequency and situs of other biocatalyst cutting sites, and the gC gene of all strains had apparente preference to G and C. There was resemblance in proteo-hydrophobicity, epitope and higher structure prediction among those strains, but also had some changes and difference.