Ⅰ群禽腺病毒4型陕西分离株33K、Fiber-1、Fiber-2、pⅢa基因的克隆及序列分析
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摘要
为明确Ⅰ群禽腺病毒陕西分离株相关基因的特征,通过聚合酶链反应(PCR)技术分别扩增了禽腺病毒陕西分离株SX17株的4个基因片段,即33K、Fiber-1、Fiber-2、pⅢa,将其分别克隆到pCold-SUMO载体后进行序列测定,并将克隆得到的4个基因片段与发表的相应序列进行序列比对及遗传进化分析。结果表明,克隆得到的4个片段与Ⅰ群禽腺病毒4型同源性最高,33K基因核苷酸及推导的氨基酸同源性在99.3%~100%与91.5%~100%之间,Fiber-1基因核苷酸及推导的氨基酸同源性在98.5%~100%与65.9%~100%之间,Fiber-2基因核苷酸及推导的氨基酸同源性在98.1%~100%与99.3%~100%之间,pⅢa基因核苷酸及推导的氨基酸同源性在98.6%~100%与98.5%~100%之间。由遗传进化树可知,这些基因在遗传关系上与血清4型位于同一分支,亲缘关系最近,分析显示陕西分离株为Ⅰ群禽腺病毒血清4型。
In order to identify the characteristics of related genes of FAV-Ⅰisolated from Shaanxi province,polymerase chain reaction(PCR)was used to amplify the four genes of FAV-Ⅰ.The gene fragments were then cloned into pCold-SUMO vector and sequenced.The results showed that 33 K gene shared 99.3%-100% nucleotide identity and 91.5%-100% amino acid identity with FAV-Ⅰ type 4.Fiber-1 gene shared 98.5%-100% nucleotide identity and 65.9%-100% amino acid identity with FAV-Ⅰ type 4.Fiber-2 gene shared 98.1%-100% nucleotide identity and 99.3%-100% amino acid identity with FAV-Ⅰ type 4.pⅢa gene shared 98.6%-100% nucleotide identity and 98.5%-100% amino acid identity with FAV-Ⅰ type 4.Phylogenetic analysis revealed that these genes seemed to be closely related to serotype 4.
In order to identify the characteristics of related genes of FAV-Ⅰisolated from Shaanxi province,polymerase chain reaction(PCR)was used to amplify the four genes of FAV-Ⅰ.The gene fragments were then cloned into pCold-SUMO vector and sequenced.The results showed that 33 K gene shared 99.3%-100% nucleotide identity and 91.5%-100% amino acid identity with FAV-Ⅰ type 4.Fiber-1 gene shared 98.5%-100% nucleotide identity and 65.9%-100% amino acid identity with FAV-Ⅰ type 4.Fiber-2 gene shared 98.1%-100% nucleotide identity and 99.3%-100% amino acid identity with FAV-Ⅰ type 4.pⅢa gene shared 98.6%-100% nucleotide identity and 98.5%-100% amino acid identity with FAV-Ⅰ type 4.Phylogenetic analysis revealed that these genes seemed to be closely related to serotype 4.
引文
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[2]罗思思,谢芝勋,邓显文,等.Ⅰ群禽腺病毒五邻体基因的克隆及原核表达[J].广东农业科学,2011,38(7):154-157.
[3]McFerran J B,Connor T J,Adair B M.Studies on the antigenic relationship between and isolate(127)from the egg drop syndrome 1976and a fowl adenovirus[J].Avian Pathol J,1978,7(4):629-636.
[4]张明明,秦爱建,庄骁颖,等.CAG和MLP启动子控制的重组禽腺病毒构建[J].畜牧与兽医,2010,42(3):23-27.
[5]Fessler S P,Young C S H.The role of the L4 33Kgene in adenovirus infection[J].Virology,1999,263(2):507-516.
[6]Gelderblom H,Maichlelauppe I.The fibers of fowl adenoviruses[J].Arch Virol,1982,72(4):289-298.
[7]Morin N,Boulanger P.Morphogenesis of human adenovirus type 2:sequence of entry of proteins into previral and viral particles[J].Virology,1984,136(1):153-167.
[8]Jackie P,Wright P J,Sheppard M.A single gene encoding the fiber is responsible for variations in virulence in the fowl adenoviruses[J].J Virol,1996,70(8):5115-5122.
[9]李茂祥,李红卫,殷震,等.减蛋综合征病毒33K蛋白基因克隆及结构分析[J].中国兽医学报,1999,19(3):221-225.
[10]文艳玲,谢芝勋,黄琦,等.禽腺病毒1型和4型六邻体蛋白抗原表位和密码子偏爱性分析[J].动物医学进展,2007,28(11):17-20.
[11]Farley D C,Brown J L,Leppard K N.Activation of the earlylate switch in adenovirus type 5major late transcription unit expression by L4gene products[J].J Virol,2004,78(4):1782-1791.
[12]武敏.Ⅰ群禽腺病毒JS株Ⅲa蛋白的原核表达及其单克隆抗体的制备[D].江苏扬州:扬州大学,2009.
[13]Kim J N,Byun S H,Kim M J,et al.Outbreaks of hydropericardium syndrome and molecular characterization of Korean fowl adenoviral isolates[J].Avian Dis,2008,52(3):526-530.
[14]Zhao J,Zhong Q,Zhang G Z,et al.Pathogenicity and complete genome characterization of fowl adenoviruses isolated from chickens associated with inclusion body hepatitis and hydropericardium syndrome in China[J].PLoS One,2015,10(7):e0133073.
[15]Li L,Luo L,Wen G,et al.Genome sequence of a fowl adenovirus serotype 4strain lethal to chickens,isolated from China[J].Genome Announce,2016,4(2):e00140-16.
[16]Asthana M,Chandra R,Kumar R.Hydropericardium syndrome:current state and future developments[J].Arch Virol,2013,158(5):921-931.
[17]袁万哲,李玉保,王建昌,等.鸡心包积液-肝炎综合征的初步研究[J].中国兽医科学,2016,46(2):157-160.
[18]袁万哲.科学防控鸡心包积液-肝炎综合征[J].中国动物保健,2016,18(2):35-35.
[19]殷震,刘景华.动物病毒学[M].2版.北京:科学出版社,1997.
[20]Tan P K,Bergelson J C M,Michou A I,et al.Defining CAR as a cellular receptor for the avian adenovirus CELO using agenetia analysis of the two viral fiber proteins[J].J Gen Virol,2001,82(6):1465-1472.