广东省鸡传染性贫血病毒编码区基因的克隆与分子进化分析
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摘要
为了解鸡传染性贫血病毒(chicken anemia virus,CAV)广东分离株变异情况,通过PCR方法克隆了于2012年分离到的10株CAV的编码区序列,并对其进行了测序及分子进化分析。结果显示,广东CAV分离株编码区全长1 823bp,含有3个互相重叠的开放阅读框(ORF)。序列分析表明,广东省CAV分离株与GenBank上已发表的其他36株CAV比较,VP1核苷酸的同源性在94.1%~99.2%之间,推导出的氨基酸的同源性在95.3%~99.3%之间;VP2的核苷酸同源性在98.2%~100%之间,推导出的氨基酸的同源性在95.9%~99.5%之间;VP3的核苷酸同源性在97.8%~100%之间,推导出的氨基酸的同源性在95.1%~99.2%之间;VP1蛋白共有17个位点发生了变异,VP2蛋白共有11个位点发生了变异,VP3蛋白共有6个位点发生了变异。分子进化分析显示,10株CAV广东分离株主要位于两个分支,其中9株CAV分离株与GenBank上已发表的35株CAV处于一个大分支,而GDN-12与分离株KC414026处于另外一个小分支,由此得出其中9株广东CAV分离株是目前主要的流行毒株。
In order to understand the variation of chicken anemia virus(CAV)strains isolated from Guangdong,the coding sequences of the Guangdong strains isolated in 2012were cloned through polymerase chain reaction(PCR)and molecular evolution of the coding region were analyzed.The coding sequences are made of 1 823bp including three overlapping open reading frames(ORF). The results of coding sequences analyses indicated that compared to 36other published CAV sequences in GenBank,the homologies of the VP1nucleotide sequences and the deduced amino acid sequences are 94.1%-99.2%and 95.3%-99.3%,respectively;the homologies of the VP2nucleotide sequences and the deduced amino acid sequences are 98.2%-100% and 95.9%-99.5%,respectively;the homologies of the VP3nucleotide sequences and the deduced amino acid sequences are 97.8%-100%and 95.1%-99.2%,respectively;the VP1protein has 17variations,the VP2protein has 11variations,the VP3protein has 6variations.Molecular evolution analysis of the complete coding sequences indicated that the isolates of Guangdong were located in two clusters,nine CAV isolates are located in the bigger group with 35other published CAV isolates in GenBank,and one CAV isolate is located in other small group with KC414026isolate.
In order to understand the variation of chicken anemia virus(CAV)strains isolated from Guangdong,the coding sequences of the Guangdong strains isolated in 2012were cloned through polymerase chain reaction(PCR)and molecular evolution of the coding region were analyzed.The coding sequences are made of 1 823bp including three overlapping open reading frames(ORF). The results of coding sequences analyses indicated that compared to 36other published CAV sequences in GenBank,the homologies of the VP1nucleotide sequences and the deduced amino acid sequences are 94.1%-99.2%and 95.3%-99.3%,respectively;the homologies of the VP2nucleotide sequences and the deduced amino acid sequences are 98.2%-100% and 95.9%-99.5%,respectively;the homologies of the VP3nucleotide sequences and the deduced amino acid sequences are 97.8%-100%and 95.1%-99.2%,respectively;the VP1protein has 17variations,the VP2protein has 11variations,the VP3protein has 6variations.Molecular evolution analysis of the complete coding sequences indicated that the isolates of Guangdong were located in two clusters,nine CAV isolates are located in the bigger group with 35other published CAV isolates in GenBank,and one CAV isolate is located in other small group with KC414026isolate.
引文
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[9]Yuasa N.Propagation and infectivity titration of the Gifu-1strain of chicken anemia agent in a cell line(MDCC-MSB1)derived from Marek’s disease lymphoma[J].Natl Inst Anim Health Q(Tokyo),1983,23(1):13-20.
[10]李延鹏,崔治中.一株鸡传染性贫血病毒野毒株致病性及其全基因组序列比较[J].微生物学报,2007,27(50):894-898.
[11]何成强,丁乃峥,李景鹏,等.鸡贫血病毒哈尔滨分离株全基因克隆和序列分析[J].微生物学报,2002,42(4):436-441.
[12]McNulty M S.Chicken anaemia agent:a review[J].Avian Pathol,1991,20(2):187-203.
[13]Otaaki Y,Nunoya T,Tajima M,et al.Depression of vaccinal immunity to Marek’s disease by infection with chicken anaemia agent[J].Avian Pathol,1998,17(2):333-347.
[2]Natesan S,Kataria J M,Dhama K,et al.Biological and molecular characterization of chicken anaemia virus isolates of Indian origin[J].Virus Research,2006,118(1/2):78-86.
[3]Kim H R,Kwon Y K,Bae Y C,et al.Molecular characterization of chicken infectious anemia viruses detected from breeder and broiler chickens in South Korea[J].Poult Sci,2010,89(11):2426-2431.
[4]崔现兰,辛桂香,呈东来,等.鸡传染性贫血病毒的鉴定[J].中国畜禽传染病,1992(6):3-5.
[5]Zhou W P,Yang B,Shen B.A serologic survey of antibody against chicken infectious anemia virus by indirect immunofluorescent assay in domestic poultry in China[J].Avian Dis,1996,40(2):358-360.
[6]Schat K A.Chicken anemia virus[J].Curr Top Microbiol Immunol,2009,331:151-183.
[7]Todd D,Connor T J,Calvert V M,et al.Molecular cloning of an attenuated chicken aneamia virus isolate following repeated cell culture passage[J].Avian Pathol,1995,24(1):171-187.
[8]Phenix K V,Meehan B M,Todd D,et al.Transcriptional analysis and genome expression of chicken aneamia virus[J].J Gen Virol,1994,75(4):905-909.
[9]Yuasa N.Propagation and infectivity titration of the Gifu-1strain of chicken anemia agent in a cell line(MDCC-MSB1)derived from Marek’s disease lymphoma[J].Natl Inst Anim Health Q(Tokyo),1983,23(1):13-20.
[10]李延鹏,崔治中.一株鸡传染性贫血病毒野毒株致病性及其全基因组序列比较[J].微生物学报,2007,27(50):894-898.
[11]何成强,丁乃峥,李景鹏,等.鸡贫血病毒哈尔滨分离株全基因克隆和序列分析[J].微生物学报,2002,42(4):436-441.
[12]McNulty M S.Chicken anaemia agent:a review[J].Avian Pathol,1991,20(2):187-203.
[13]Otaaki Y,Nunoya T,Tajima M,et al.Depression of vaccinal immunity to Marek’s disease by infection with chicken anaemia agent[J].Avian Pathol,1998,17(2):333-347.