1.Gene clone, eukaryotic expression vector and tissue expression profile analysis of porcine BST-2 gene
Ning KONG ; Yongguang WU ; Qiong MENG ; Zhongze WANG ; Wu TONG ; Hao ZHENG ; Guoxin LI ; Tongling SHAN ; Enmin ZHOU ; Guangzhi TONG
Chinese Journal of Veterinary Science 2017;37(8):1594-1599,1640
In order to study the biological function of pig BST-2 gene,the BST-2 gene was amplified with specific primers from porcine kidney tissue,and molecular characterization of BST-2 nuclectide and amino acid sequence were analyzed with bioinformatics tools and online server.Then the prokaryotic expression and tissue expression profile analysis was carried out.The results showed that the full length of pig BST-2 gene was 851 bp and contained 23 bp of 5'-UTR,294 bp of 3'-UTR and 534 bp of CDS and the gene encoded 177 aa.Amino acid sequence analysis of pig BST-2 protein showed 46.1% identity with gorilla gorilla,41.7% with cricetulus griseus,39.5% with mus musculus,35.4% with equus asinus,42.0% with felis catus,40.5% with bos mutus,44.4% with macaca mulatta,38.7% with ovis aries and 46.8% with homo sapiens.BST-2 protein contained 2 transmembrane structure (27-49 aa and 154-176 aa),2 glycosylation sites and 14 potential phosphorylation sites including ATM,CK Ⅱ,PKA,PKC binding sites.The pig BST-2 protein was expressed in Vero cells after translated the recombinant plasmid FLAG-BST-2.Semiquantitative PCR results showed that BST-2 gene was expressed in all the tissues,especially in lymph nodes,thymus,tonsils,spleen,large intestine and small intestine.This study provide a foundation for further understanding the antiviral mechanism of pig BST-2 protein.
2.Molecular characterization of Japanese encephalitis virus strains prevalent in Chinese swine herds.
Hao ZHENG ; Tongling SHAN ; Yu DENG ; Chunqing SUN ; Shishan YUAN ; Yang YIN ; Guangzhi TONG
Journal of Veterinary Science 2013;14(1):27-36
Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals
;
Cell Line
;
Cricetinae
;
Encephalitis Virus, Japanese/classification/*genetics
;
Encephalitis, Japanese/epidemiology/*veterinary/virology
;
Gene Expression Regulation, Viral/physiology
;
Genome, Viral
;
Molecular Epidemiology
;
Phylogeny
;
Swine
;
Swine Diseases/epidemiology/*virology
3.Molecular characterization of Japanese encephalitis virus strains prevalent in Chinese swine herds.
Hao ZHENG ; Tongling SHAN ; Yu DENG ; Chunqing SUN ; Shishan YUAN ; Yang YIN ; Guangzhi TONG
Journal of Veterinary Science 2013;14(1):27-36
Japanese encephalitis virus (JEV) is the leading cause of viral encephalitis in Asia and domestic pigs serve as the amplifying hosts. In the present study, the full genomic sequences of two JEV strains (HEN0701 and SH0601) isolated from pigs in China were determined and compared with other 12 JEV strains deposited in GenBank. These two strains had an 88.8% nucleotide sequence similarity and 97.9% deduced amino acid sequence homology. HEN0701 had high nucleotide sequence and high amino acid sequence identity with genotype I (GI) strains, while SH0601 had high nucleotide sequence and high amino acid sequence identity with GIII strains at both the gene and full genome levels. Further phylogenetic analysis showed that HEN0701 belonged to the JEV GI group and SH0601 was classified as a GIII strain. Analysis of codon usage showed there were a few differences between the GI and GIII strains in nucleotide composition and codon usage for the open reading frames.
Animals
;
Cell Line
;
Cricetinae
;
Encephalitis Virus, Japanese/classification/*genetics
;
Encephalitis, Japanese/epidemiology/*veterinary/virology
;
Gene Expression Regulation, Viral/physiology
;
Genome, Viral
;
Molecular Epidemiology
;
Phylogeny
;
Swine
;
Swine Diseases/epidemiology/*virology