1.Dual Expression of Two Transgenes Introduced by Lentiviral Vectors.
Su Jung PARK ; Sun Ju CHOI ; Joo Young PARK ; Kyoung Ho LEE
Journal of Bacteriology and Virology 2005;35(2):157-164
Lentiviruses can infect mitotic and non-dividing cells owing to the karyophilic properties of their pre-integrating complex, which allow its active import through the nucleopore. Thus lentiviral vectors derived from human immunodeficiency virus type 1 can mediate an efficient transfer integration, and stable expression of transgenes into proliferating and stationary cells both in vivo and in vitro. By adopting the internal ribosome entry site of encephalomyocarditis virus for bicistronic expression or two promoters of EF-1alpha and SV40 for separate expression of two genes of interest, we developed two lentiviral vectors that express two genes. On FACS analysis, RT-PCR, and immunofluorescence assay, it was shown that the target cells expressed two genes of interest at different levels as the transducing vectors designed for. This vector system is useful especially for a stable, dual-gene expression and two transgene deliveries to non-dividing cells.
Encephalomyocarditis virus
;
Fluorescent Antibody Technique
;
HIV-1
;
Lentivirus
;
Peptide Elongation Factor 1
;
Ribosomes
;
Transgenes*
2.Isolation, identification and full-length genome sequence analysis of encephalomyocarditis virus from local aardvarks.
Hong-Tao CHANG ; Hui-Min LIU ; Xiu-Yuan HE ; Jun ZHAO ; Lu CHEN ; Xin-Wei WANG ; Xia YANG ; Hui-Xia YAO ; Chuan-Qing WANG
Chinese Journal of Virology 2014;30(4):375-381
Encephalomyocarditis virus (EMCV) is a natural epidemic zoonotic pathogen. However, no reports have been published regarding the isolation, identification and full-length genome of EMCV from a local aardvark population. In present study, an EMCV isolate HNXX13 was isolated from aardvarks named Huainan-pig in Henan Province. The systematic identification, full-length genome sequencing and molecular characteristic analysis of the isolate HNXX13 were conducted. The result showed that the isolate was spherical with a diameter of 24-30 nm, neither heat- nor acid-resistant, sensitive to trypsin, insensitive to chloroform, not protected by bivalent cationic, and the specific fluorescence was observed in the cytoplasm of BHK-21 cells infected with the isolate by using indirect fluorescence assay. The full-length genome of EMCV HNXX13 generated a 7 725bp sequence (GenBank: F771002), with 81.0%-99.9% nucleotide identity to reference strains from different animals, and 99.5% with a Chinese reference strain isolated earlier from a commercial pig herd. The phylogenetic tree based on the full-length genome and ORF sequences identified that all EMCV strains were divided into three groups G1, G2 and G3, and strain HNXX13 belonging to the G1 group with other Chinese reference strains. The result also identified that this EMCV infection could cause severe clinical signs in a local aardvark population, and enriches the molecular epidemiological data of EMCV in China. Regional differences exist in EMCV genome and transmission is limited within a certain area. However, the cross-infection and transmission of EMCV between aardvark and mice appears most likely. Mutations have occurred in some amino acids of EMCV strain HNXX13 during the transmission in local aardvark herd and these mutations might make the virus easier to infect the aardvark.
Animals
;
Animals, Wild
;
virology
;
Cardiovirus Infections
;
veterinary
;
virology
;
China
;
Encephalomyocarditis virus
;
classification
;
genetics
;
isolation & purification
;
Genome, Viral
;
Mice
;
Molecular Sequence Data
;
Phylogeny
;
Xenarthra
;
virology
3.Sequencing and analysis of the complete genome of encephalomyocarditis virus strain GXLC isolated from swine.
Kai-Chuang SHI ; Su-Jie QU ; Jin-Xi CHEN ; Rui-Sheng XU ; Min ZHENG ; Qi LIU ; Han-Zhong CHEN ; Gang LI
Chinese Journal of Virology 2010;26(2):134-142
The complete genome of encephalomyocarditis virus (EMCV)strain GXLC isolated from swine was sequenced and analyzed. Five overlapped gene fragments covering the entire open reading frame (ORF) were amplified by RT-PCR, and the 3'-untranslated region (UTR) and 5'-UTR were amplified by the 3'-rapid amplification of cDNA ends (RACE) and 5'-RACE method, respectively. The genome sequences of strain GXLC were obtained by assembling the sequences of RT-PCR-generated cDNA fragments. The length of the complete genome was 7 725 nucleotides (nt). The homology comparison and phylogenetic analysis of the nucleotide and deduced amino acid sequences between strain GXLC and other EMCV strains available in GenBank were performed. The results showed that the complete genome identity between GXLC strain and the strains from China, i.e. GX0601, GX0602, BJC3 and HB1 and the strains from other countries, i.e. CBNU, K3, K11, TEL-2887A, EMCV-R and PV21 was over 99%. The phylogenetic trees based on the complete genome, the structural protein or the non-structural protein gene sequences revealed that the tree topology was similar. All the EMCV strains could be divided into two groups: group I and group II, and group I could be subdivided into subgroup Ia and subgroup Ib. The strains from swine belonged to subgroup Ia or Ib, and the strains from mice belonged to subgroup Ia, while the strains from Sus scro fa belonged to group II. Strain GXLC, together with other EMCV isolates from China, belonged to subgroup Ia.
Animals
;
Cardiovirus Infections
;
veterinary
;
virology
;
Cell Line
;
Encephalomyocarditis virus
;
classification
;
genetics
;
isolation & purification
;
Genome, Viral
;
Molecular Sequence Data
;
Open Reading Frames
;
genetics
;
Phylogeny
;
Reverse Transcriptase Polymerase Chain Reaction
;
Sequence Analysis, DNA
;
Species Specificity
;
Swine
;
Swine Diseases
;
virology
;
Weaning