Molecular Cloning And Bioinformatics Analysis Of Pseudorabies Virus Protein Kinase Gene

There are 12 pseudorabies virus strain,SS,SQ and Fa,783strain,Bartha,(purchased from China Institue of Veterinary Drug Control) and SA215(purchased from China Animal Husbandry Industry CO.LTD)and SN,SL,SCZ(Isolated by Xu Zhiwen associate professor in Sichuan Agriculture University Animal biotechnology center in2000) and BJ,DG,HS(gived by Lou Gaoming professor in Shaoguan University College of Yingdong Biomgineering).Designing a pair of primer according to PRV PK gene,we received 12 PK complete gene by PCR.We used TaKaRa's kit to retrieve gel and to ligate, transform into E.coli DH_5a,and picking singal white masculine masculine by Amp-X-gal-IPTG LB flat plate.After identified by COL-PCR and RE,they were sequenced by TaKaRa and GeneCore.12 sequenced PK genes of different PRV strains,as well as 6 PK genes which downloaded from GenBank were analyzed by bioinformatics software,the analysis and prediction of these genes including nucleotide homology,codons bias,location of mutation area,phylogenetic tree,amino acids homology,protein hydrophilicity and epitope.At the same time,analyzing the nucleotide homology,amino acids homology and the protein domain of PK genes of Herpesvirus.The results showed that the ORF length of PRV-PK gene is 1107-1164nt,amino acids length is 369-388,nucleotide homology is 96.7%-100%, and amino acids homology is 73.9%-100%.Two hypermutation replicated plot is located in 1079～1099nt and 240～250nt.Different strains were prefer to GC basi-.The results of different strains of enzyme site,protein hydrophilicity and epitope were very similar.The nucleotide homology and amino acids of Herpesvirus is 2.2%-51.5%and 8.2%～39.3%.The amino acids of PRV PK contain most of the conserved motifs of a eukaryotic serine/threonine protein kinase.The PK among Herpesvirus has some featured common amino acids sequence.