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| 马传染性贫血病毒强毒株gag和gp45基因的体内进化分析 | |
| 引用本文： | 刘强,王雪峰,王珊珊,韦华冕,杜承,林跃智,马建,周建华,王凤龙.马传染性贫血病毒强毒株gag和gp45基因的体内进化分析[J].动物医学进展,2012,33(7):1-7. |
| 作者姓名： | 刘强 王雪峰 王珊珊 韦华冕 杜承 林跃智 马建 周建华 王凤龙 |
| 作者单位： | 1. 内蒙古农业大学兽医学院,内蒙古呼和浩特010018;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江哈尔滨150001 2. 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江哈尔滨,150001 3. 内蒙古农业大学兽医学院,内蒙古呼和浩特,010018 |
| 基金项目： | 国家自然科学青年基金项目，国家"十二"五重大传染病专项 |
| 摘 要： | 为研究马传染性贫血病毒(EIAV)强毒株体内进化规律,将3匹马分别以1×105 TCID50/匹接种EIAVLN40强毒株后,在不同时间点采取外周血,分离单核白细胞。提取白细胞中的DNA,并以其为模板,应用套式PCR技术分别扩增EIAV前病毒DNA的p15,p9和gp45基因,经克隆后进行序列分析。结果显示,与接种前EIAVLN40相比较,p15基因核苷酸和氨基酸平均差异率分别为1.8%(0～2.9%)和1.7%(0～3.6%);p9分别为2.5%(0.3%～3.8%)和2.9%(0～4.8%);gp45-Ⅰ分别为1.3%(0.3%～2.1%)和1.8%(0～3.2%),gp45-Ⅱ分别为2.1%(0～3.4%)和2.6%(0～4.7%)。在EIA发热期,每个基因核苷酸和氨基酸差异率最小;在亚临床阶段,每个基因平均差异率总体上相对较高。氨基酸变异位点分析发现,发病期(2-2、3-1)各基因氨基酸序列回复到亲本株EIAVLN40,其余时期p15基因氨基酸稳定变异位点16N/S、88T/S和112M/E;p9基因115E/G和122K/M;gp45-Ⅰ基因9T/F/L;gp45-Ⅱ基因21E/K和29G/S。此外,p15基因进化树结果显示,发热期(2-2、3-1)p15基因序列与EIAVLN40序列处于同一分支上,亚临床阶段大多数p15基因序列处于另一个大分支上;其他基因进化情况与p15基因类似。对EIAV p15、p9、gp45基因体内进化分析,有助于对EIAV在马体内感染进程的研究,为进一步研究EIAV持续感染和逃避免疫监视的机制提供参考。 |
| 关 键 词： | 马传染性贫血病毒 p15基因 p9基因 gp45基因 差异率 变异分析 |
In Vivo Evolution of gag and gp45 Genes of Equine Infectious Anemia Virus |
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| LIU Qiang , WANG Xue-feng , WANG Shan-shan , WEI Hua-ming , DU Cheng , LIN Yue-zhi , MA Jian , ZHOU Jian-hua , WANG Feng-long.In Vivo Evolution of gag and gp45 Genes of Equine Infectious Anemia Virus[J].Progress In Veterinary Medicine,2012,33(7):1-7. | |
| Authors: | LIU Qiang WANG Xue-feng WANG Shan-shan WEI Hua-ming DU Cheng LIN Yue-zhi MA Jian ZHOU Jian-hua WANG Feng-long |
| Institution: | 1(1.College of Veterinary Science,Inner Mongolia Agricultural University,Huhhot,Inner Mongolia,010018,China; 2.State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin,Heilongjiang,150001,China) |
| Abstract: | To study the evolution rule of the EIAVLN40 in vivo,3 horses were inoculated with EIAVLN401×105TCID50 respectively,the peripheral blood lymphocytes(PBL) were collected from blood at different time points.DNA extracted from PBL were used as the template strand to amplify the EIAV proviral DNA of p15,p9 and gp45 region by nested PCR technique.The cloning,identification and sequencing were conducted.The result showed that the average divergences of nucleotide and amino acid of p15 gene were 1.8%(0-2.9%) and 1.7%(0-3.6%) respectively,p9 were 2.5%(0.3%-3.8%)and 2.9%(0-4.8%)respectively,gp45-Ⅰwere 1.3%(0.3-2.1%)and 1.8%(0-3.2%)respectively,gp45-Ⅱwere 2.1%(0-3.4%) and 2.6%(0-4.7%)respectively,compared with EIAVLN40.Moreover,during the fever period of EIA,the divergence of each nucleotide and amino acid was in a lower level or even no change;While the rate was relatively high in the sub-clinical stage.Analysis on amino acids variation found that during the fever period of EIA,amino acid sequence of each gene(2-2,3-1) reverts to the field strain EIAVLN40;in the sub-clinical stage,substitutions of translated amino acid residues of p15 gene were in the 16th N/S,88th T/S and 122nd M/E about three horses;p9 gene was in the 115th E/G and 122nd K/M about three horses;gp45-Ⅰgene was in the ninth T/F/L about three horses;gp45-Ⅱgene was in the 21st E/K and 29thG/S about three horses.In addition,phylogenetic tree analysis of p15 gene(2-2,3-1) showed that the gene sequences in fever period were on the same branch with EIAVLN40,but others at other time points were on another branch;Evolutionary analysis of other genes is similar to the p15 gene.Our data on the EIAV p15,p9 and gp45 evolution in vivo facilitate the studies on procedures of infection of EIAV pathogeny.The result would provide a reference to further study EIAV in persistent infection and the mechanism of evading immune surveillance. |
| Keywords: | Equine infectious anemia virus p15gene p9gene gp45gene divergence variation analysis |
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