| 猪巨细胞病毒的PCR检测及其gB基因特征分析 |
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| 引用本文： | 李晶,余兴龙,李润成,罗维,向卫军,李薇,王亚,葛猛.猪巨细胞病毒的PCR检测及其gB基因特征分析[J].湖南农业大学学报(自然科学版),2009,35(5). |
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| 作者姓名： | 李晶 余兴龙 李润成 罗维 向卫军 李薇 王亚 葛猛 |
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| 作者单位： | 湖南农业大学,动物医学院,湖南,长沙,410128 |
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| 基金项目： | 湖南省科技重大专项，湖南农业大学人才引进基金(04YJ1030 |
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| 摘 要： | 根据GenBank登录的猪巨细胞病毒(PCMV)gB基因序列设计检测引物和全长gB基因引物,对来自湖南浏阳和江西景德镇不同养殖场的猪鼻拭子样品(51份)进行PCR检测,60.78%(31/51)的鼻拭子样品扩增出了260 bp的特异性条带.再以阳性样品为模板扩增全长gB基因,并将其克隆到pMD18-T载体,核苷酸序列测定和结果分析表明,所获得PCMVgB基因序列全长2 580 bp,编码860个氨基酸,与国外PCMV分离株的核苷酸同源性为97.6% ～ 98.9%,氨基酸同源性为97.0%～98.6%,且 PCMV可分为2个群:一群为中国湖南株、英国株和德国株,命名为A群;另一群为日本株、瑞士株和西班牙株,命名为B群.抗原表位优势、亲水性、表面可及性预测分析表明,PCMVgB蛋白是理想的免疫学抗原候选蛋白.
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| 关 键 词： | 猪巨细胞病毒 gB基因 序列分析 |
PCR detection and sequence analysis of gB gene of porcine cytomegalovirus |
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| Abstract: | According to the gB gene of porcine cytomegalovirus (PCMV) from GenBank, two pairs of primers were designed and used for polymerase chain reaction (PCR) and diagnoses of clinical samples with nasal swab specimens. Two specific DNA fragments were amplified, and gB gene was cloned into PMD18-T vector. The recombinant plasmids were sequenced and compared with published sequence. The results showed that 31 of 51 clinical samples from different regions in Hunan province and Jiangxi province were positive, with a positive rate of 60.78%. Analysis of sequence showed that the PCMV gene was 2580 nt in size and encoding 860 AA. In comparison with sequences of 5 PCMV isolates in the world registered in GenBank the PCMVgB strain, they shared 97.6%~98.9% nucleotide sequence homology, and 97.0%~98.6% amino acid sequence homology, respectively. According to the phylogenetic tree of glycoprotein B gene nucleotide, PCMV can be divided into groups A and B. Group A includes China HN strain, England strain and German strain; Group B includes Japan strain, Sweden strain and Spain strain. Analysis of antigentic index, hydrophilicity plot and surface probability of gB protein of PCMV indicate that it is an ideal candidate protein antigen in immunology. |
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| Keywords: | PCR |
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