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Expression of Porcine Circovirus Type 2 orf2 gene in Myocardial Cells of Detached Cherry Valley Duck

Agricultural Biotechnology 2014年第5期3卷 51-54页

作者： Wenxiu WANG Sufang LU Shijun FU Guangjun GUO Ling MO Yueqiang XIAO Zhiqiang SHEN Shandong Binzhou Animal Science&Veterinary Medicine Academy Shandong Lvdu Bio-Sciences &Technology Co. Ltd.

The objective of this study was to construct the recombinant eukaryotic expression plasmid of orf2 gene harboring enhanced green fluorescent protein （EGFP） report gene. orf2 gene of porcine circovirus type 2 cloned by PCR was ligated to the expression vector pEGFP-N1, which contains enhanced green fluorescent protein （EGFP） report gene, the recombinant eukaryotic expression plasmid pEGFP-N1-orf2 was constructed successfully and was transfected into pre- pared duck myocardial cells （DMCs） by lipofectin. According to the result, the fluorescence expression was directly detected with fluorescence microscope, and the expression of orf2 were analyzed by RT-PCR and indirect immunofluorescence assay （IFA） respectively. About 48 h after transfection, green fluorescent can be observed on transfected cells ： T-PCR and IFA were positive. This indicated that orf2 gene of PCV2 was expressed efficiently in transfected duck myocardial cells.

关键词： Porcine circovirus orf2 gene Duck myocardial cells Expression

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Cloning and Sequence Analysis of orf2 gene of Seven Porcine Circovirus Type 2

Cloning and Sequence Analysis of ORF2 Gene of Seven Porcine ...

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第五届中国畜牧科技论坛

作者： L Su-fang WANG Wen-xiu GUO Guang-jun WEI Feng TANG Na GUAN Yu SHEN Zhi-qiang Shandong Binzhou Animal Science & Veterinary Medicine Insitute

In order to research the orf2 genes of separate PCV2,the orf2 gene was amplified from seven tissues of piglets suffered from PMWS,respectively using the primers designed according to the orf2 sequences published in GenBank.The resultant PCR products were cloned into pMD18-T vectors.The recombinant plasmids respectively named pMD-PCV2-HN01,pMD-PCV2-DY01,pMD-PCV2-HM01,pMD-PCV2- SDqd01,pMD-PCV2-SDqd02,pMD-PCV2-SDqd03 and pMD-PCV2-PD01 were sequenced and analysed. The results indicated that the orf2 gene of HN01 isolate was 705bp,while others were 702bp.The nucleotide homology among seven genes were 94.0%to 99.9%and amino acid homology were high than 93.6%.The nucleotide homologies between the seven isolates and German isolate or Taiwan isolate in GenBank were lower from 91.7%to 93.2%and 89.0%to 90.5%,whereas,that were higher than 94.0%between the seven isolates and Denmark,France or China isolates,amino acid homology were high than 89.3%.

关键词： porcine circovirus type 2 orf2 gene cloning sequence analysis

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Isolation and phylogenetic analysis of feline calicivirus strains from various region of China

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Animal Diseases 2022年第3期2卷 165-174页

作者： Longlong Cao Qiuyan Li Kaituo Shi Liting Wei Hehao Ouyang Zijun Ye Wenguang Du Jiawen Ye Xiaochen Hui Jiakang Li Shengbo Cao Dengyuan Zhou State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhanChina College of Veterinary Medicine Huazhong Agricultural UniversityWuhanChina Wuhan Keqian Biology Co. LtdWuhanChina

Feline calicivirus(FCV)is an important feline pathogen mainly causing upper respiratory tract disease,conjunctivitis,and stomatitis,and it is classifed into genotype I and genotype II.To investigate the prevalence and molecular characteristics of FCV,this study collected 337 cat swab samples from animal hospitals in diferent regions of China from 2019 to 2021.The positive detection rate of FCV was 29.9%(101/337)by RT-PCR.Statistical analysis showed that FCV prevalence was signifcantly associated with living environment(p=0.0004),age(p=0.031)and clinical symptoms(p=0.00),but not with sex(p=0.092)and breed(p=0.171).The 26 strains of FCV were isolated using F81 cells.Phylogenetic analysis showed that 10 isolates belonged to genotype I,and 16 isolates belonged to genotype II.These 26 isolates were highly genetically diverse,of which HB7 isolate had three same virulence-related amino acid loci with VSD strains.Potential loci distinguishing diferent genotypes were identifed from 26 isolates,suggesting the genetic relationship between diferent genotypes.In addition,selection pressure analysis based on capsid protein of 26 isolates revealed that the protein is under diversifying selection.This study reveals the genetic diversity of FCV and provides a reference for the screening of vaccine candidate strains and the development of vaccines with better cross-protection efects.

关键词： Feline Calicivirus genetic diversity Phylogenetic analysis Selection pressure analysis orf2 gene

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