| 鸡肾型IBV分离株M基因的克隆与特性分析 |
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| 引用本文： | 王晶钰,罗艳,邓小敏,郭抗抗,张彦明.鸡肾型IBV分离株M基因的克隆与特性分析[J].中国预防兽医学报,2007,29(8):591-595. |
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| 作者姓名： | 王晶钰 罗艳 邓小敏 郭抗抗 张彦明 |
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| 作者单位： | 西北农林科技大学,动物科技学院,陕西,杨凌,712100 |
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| 摘 要： | 依据NCBI所登录的鸡传染性支气管炎病毒的M基因序列设计了一对引物,应用TRIzol试剂盒对8个肾型鸡传染性支气管炎病毒陕西地方分离株进行总RNA的提取,以所得到的RNA为模板,用RT-PCR对M基因进行扩增;其目的条带回收提纯后,与PMD18-T克隆载体进行连接,转化到DH5α宿主菌,并经药物抗性筛选、PCR及酶切鉴定,将所筛选鉴定出的阳性质粒进行测序,最后对测序结果进行分析。结果表明：陕西地方8个毒株（BJ2、FF2、YL2、B01、G5、YX、WH、WG）M基因长约为650 bp,其中YX、WH株本身无BamH 1酶切位点。WH与其它分离株的核苷酸同源性为91.8%～92.6%,而其它的分离株间的同源性为98.8%～99.8%。8个毒株与参考株的核苷酸同源性为74.9%～99.8%。其N端90 aa肽段与对照株相比,不同之处表现为高亲水性氨基酸取代低亲水性氨基酸,这会使其具有更好的抗原性。
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| 关 键 词： | 鸡肾型IBV M基因 克隆与分析 |
| 文章编号： | 1008-0589（2007）08-0591-05 |
| 修稿时间： | 2006-10-25 |
Cloning and characterization of matrix glycoprotein gene of nephrotropic avian infectious bronchitis virus |
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| WANG Jing-yu,LUO Yan,DENG Xiao-min,GUO Kang-kang,ZHANG Yan-ming.Cloning and characterization of matrix glycoprotein gene of nephrotropic avian infectious bronchitis virus[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(8):591-595. |
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| Authors: | WANG Jing-yu LUO Yan DENG Xiao-min GUO Kang-kang ZHANG Yan-ming |
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| Institution: | College of Animal Science and Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling 712100, China |
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| Abstract: | The M genes of 8 infectious bronchitis virus(IBV)isolates from shan'xi were amplified by RT-PCR using a pair of primers designed according to the published Matrix glycoprotein gene sequence of IBV.The PCR products were cloned into PMDIS-T vector and sequenced.Analysis of the M gene sequences showed that these local isolates shared 98.8%-99.8% nucleotide homology,The WH isolate was slightly divergent and shared 91.8%-92.6% identity with the others.Comparing to reference strains,the local isolates shared 74.9%-99.8% sequence identity with the published sequences.Furthermore,the amino acids in the N-terminal 90 aa region of the local isolates were substituted by the amino acids with high hydrophilicity,indicating a better antigenity of M gene in these isolates. |
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| Keywords: | avian nephropathgenic infectious bronchitis virus M gene cloning and analysis |
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