| Two pig's abortus was resulted of pseudorabies virus by gD-PCR detected in 14 pig's abortus which were picked up from several large-scale hogpen in Sichuan province between 2004. 6-2005. 6. And then we Isolated two pseudorabies virus strain from these two pig's abortus successfully.They are named SS and SQ according to where they were picked up.There are 12 pseudorabies virus strain ,SS,SQ and Fa (saved by Sichuan Agriculture University Animal biotechnology center) and SA215 ,783strain , Bartha(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 inl998) and BJ,DG,HS (gived by Lou Gaoming professor in Shaoguan University College of Yingdong Biorngineering) . Designing a pair of primer according to PRV gD gene, we received 12 gD complete gene by PCR.We used TaKaRa' s kit to retrieve gel and to ligate, transform into E. coli DH5a, and picking singal white masculine masculine by Amp-X-gal-IPTG LB flat plate. After identified by COL-PCR and RE, they were sequenced .We sequenced 12 PRV-gD gene and downloaded 9 gD's sequence , and then predict and analyzed their's Homology of nucleotide and amino acids sequence ,lacated mutation area,phylogentic tree,Enzymatic digestion site, code preference,Hydrophobicity, Predicting Antigenic helices and Three-dimensional structure by biology software.The result is : PRV-gD's ORF's length is between 1197-1215nt, amino acids' length is between 399～405; Homology of nucleotide sequence is between 97.3%~100%, Homology of amino acids sequence is between 89.8%~98.8%;PRV's code pref GC very much; phylogentic tree analusis result is make three district where PRV spreaded in Chian: Sichuan Province ,Northern Chinaan and Southeast China; and other result is very similar. All above is demonstrate; document;evid.;evidence;justify;proof;testify; verification;witness marks PRV-gD is very important to PRV and it has a heightly conservatism.. |
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