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Molecular Cloning and Prokaryotic Expression of Non-Structural Protein NS1 Gene of Porcine Parvovirus

Agricultural Sciences in China 2003年第10期2卷 1175-1178页

作者： WUDan tongguang-zhi QIUHua-ji XUEQiang ZHOUYan-jun LIJing-peng NationalKeyLaboratoryofVeterinaryBiotechnology HarbinVeterinaryResearchInstituteHarbin150001P.R.China DepartmentofBiotechnology NortheastAgriculturalUniversityHarbin150030P.R.China

Porcine parvovirus (PPV) is one of the major agents causing swine reproductive failure. NS1 protein is a non-structural protein of PPV and can be used as a reagent for differentiation of vaccinated animals and infected ones. In present study, a recombinant plasmid pET28a/NS1 was constructed by cloning the coding sequence for NS1 of PPV into pET28a, a bacterial expression vector. The NS1 protein was expressed in E. coli BL21(DE3) after induced by IPTG and the recombinant fusion protein was purified with affinity chromatography. Expression amount of NS1 protein was improved by optimizing the inducing parameters. The recombinant NS1 protein is reactive to PPV positive sera in Western blot and ELISA test and therefore can be applicable in differential diagnosis of PPV infections.

关键词： Porcine parvovirus NS1 Diagnosis

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Immune Responses in Mice Injected with gD Plasmid DNA of Infectious Bovine Rhinotracheitis Virus

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Journal of Northeast Agricultural University(English Edition) 2004年第1期11卷 87-89页

作者： LIJi-chang tongguang-zhi QIUHua-ji AnimalMedicalCollege NortheastAgriculturalUniversityHarbinHeilongjiang150030PRC HarbinVeterinaryResearchInstitute CAASHarbinHeilongjiang150001PRC

The gene encoding gD of isolate Luojing of infectious bovine rhinotracheitis virus(IBRV)was amplified, sequenced, and cloned into plasmid pcDNA 3.1, resulting in a recombinant pcDNA-gD. Groups of BALB/c mice were injected with 100 μg of plasmid only or together with liposome. After immunization, serum samples were collected from mice every 2 weeks for a 10-week period and tested for protein-specific antibody with enzyme-linked immunosorbent assay(ELISA). It was showed that the plasmid encoding IBRV glycopretein D developed gene-specific antibody. This report indicates the potential of DNA injection as a method of vaccination.

关键词： IBRV glycoprotein D gene gene vaccination ELISA

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Cloning and Sequence Analysis of Glycoprotein D Gene of Bovine Herpesvirus-1 Strain Luojing

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Journal of Northeast Agricultural University(English Edition) 2003年第2期10卷 137-140页

作者： LIJi-chang tongguang-zhi QIUHua-Ji ZHOUYan-Jun XUEQiang NortheastAgriculturalUniversity HarbinHeilongjiang150030PRC HarbinVeterinaryResearchInstitute.Harbin Heilongjiang150001PRC

By means of PCR,the gene encoding gD of bovine herpesvirus-1 (BHV-1) strain Luojing was amplified,cloned and *** nucleotide sequence of this gD gene was (1 251 bp,)encoding 417 amino *** with the published P8-2 strain,the homology of the necleotide sequence is 99.92%,and that of the deduced amino acid sequence is 100%.The results indicated that gD of BHV-1 was highly conservative.

关键词： bovine herpesvirus-1(BHV-1) D glycoprotein gene(gD) cloning sequence analysis.

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