犬IFN-β的克隆及其序列分析
|
摘要
为了分析犬干扰素β(IFN-β)基因分子特征,预测其编码蛋白质的生物学功能,根据GenBank登录的犬IFN-β基因序列(FJ194477)设计合成特异性引物,通过PCR从外周血淋巴细胞总DNA中扩增IFN-β基因CDS区并克隆,应用生物信息学方法对犬IFN-β基因进行序列分析,并对其理化性质以及编码蛋白的二级结构、三级结构进行预测,构建分子系统进化树。结果表明:犬IFN-β基因编码区全长561 bp,共编码186个氨基酸,其分子式可表示为C_(1014)H_(1593)N_(263)O_(290)S_9,分子质量为22.40 ku;属于亲水性蛋白,理论等电点为5.98;含有丰富的二级结构,以α-螺旋(76.88%)和无规则卷曲(19.35%)为主,蛋白三级结构呈弯曲螺旋结构;与貉、威德尔海豹、金钱豹、家猫、宽吻海豚、人、鸡和绿头鸭相对应序列核苷酸序列同源性分别为98.4%、87.2%、85.2%、84.7%、79.0%、74.9%、43.9%和43.3%。说明犬与貉IFN-β核苷酸同源性最高,亲缘关系最近。
引文
[1] 曲嘉琪,李洋,钟一鸣,等.IFN-β及其研究进展[J].安徽农业科学,2013,41(9):3792-3793,3852.
[2] KLOTZ D,BAUMGARTNER W,GERHAUSER I. Type Ⅰ interferons in the pathogenesis and treatment of canine diseases[J]. Vet Immunol Immunopathol,2017,191:80-93.
[3] WOELK C H,FROST S D,RICHMAN D D,et al. Evolution of the interferon alpha gene family in eutherian mammals[J]. Gene,2007,397(1/2):38-50.
[4] YANG L,XU L,LI Y,et al. Molecular and functional characterization of canine interferon-epsilon[J]. J Interferon Cytokine Res,2013,33(12):760-768.
[5] 张华金,李娜,杜学志.干扰素的剂型及临床应用[J]. 华西药学杂志,2004,19(1):53-55.
[6] HIMMLER A,HAUPTMANN R,ADOLF G R,et al. Structure and expression in Escherichia coli of canine interferon-alpha genes[J]. J Interferon Res,1987,7(2):173-183.
[7] DERYNCK R,CONTENT J,DECLERCQ E,et al. Isolation and structure of a human fibroblast interferon gene [J]. Nature,1980,285:542-546.
[8] CAI M H,ZHU F,SHEN P. Expression and purification of chicken beta interferon and its antivirus immunological activity[J]. Protein Expr Purif,2012,84(1):123-129.
[9] 徐雷,冯若飞,李向茸,等.猪IFN-β真核表达载体pEGFP-C1-IFN-β的构建及在CHO细胞中的表达[J]. 中国兽医科学,2015,45(7):751-757.
[10] LIN F C,YOUNG H A. Interferons:success in anti-viral immunotherapy[J]. Cytokine Growth Factor Rev,2014,25(4):369-376.
[11] HENSLEY L E,FRITZ L E,JAHRLING P B,et al. Interferon-beta 1a and SARS coronavirus replication [J]. Emerg Infect Dis,2004,10 (2):317-319.
[12] KOERNER I, KOCHS G, KALINKE U, et al. Protective role of beta interferon in host defense against influenza A virus[J]. J Virol,2007,81(4):2025-2030.
[13] IWATA A,SAITO T,MIZUKOSHI-LWATA N,et al. Cloning and expression of the canine interferon-beta gene[J]. J Interferon Cytokine Res,1996,16(10):765-770.
[14] 谭明芬. 犬IFN-β的基因克隆、原核表达及活性测定[D].扬州:扬州大学,2010.
[15] 张敏,张鑫宇,夏晓莉,等.类弹性蛋白多肽融合犬IFN-β的表达纯化和抗病毒活性及体外半衰期的研究[J].中国兽医科学,2016,46(12):1543-1549.
[2] KLOTZ D,BAUMGARTNER W,GERHAUSER I. Type Ⅰ interferons in the pathogenesis and treatment of canine diseases[J]. Vet Immunol Immunopathol,2017,191:80-93.
[3] WOELK C H,FROST S D,RICHMAN D D,et al. Evolution of the interferon alpha gene family in eutherian mammals[J]. Gene,2007,397(1/2):38-50.
[4] YANG L,XU L,LI Y,et al. Molecular and functional characterization of canine interferon-epsilon[J]. J Interferon Cytokine Res,2013,33(12):760-768.
[5] 张华金,李娜,杜学志.干扰素的剂型及临床应用[J]. 华西药学杂志,2004,19(1):53-55.
[6] HIMMLER A,HAUPTMANN R,ADOLF G R,et al. Structure and expression in Escherichia coli of canine interferon-alpha genes[J]. J Interferon Res,1987,7(2):173-183.
[7] DERYNCK R,CONTENT J,DECLERCQ E,et al. Isolation and structure of a human fibroblast interferon gene [J]. Nature,1980,285:542-546.
[8] CAI M H,ZHU F,SHEN P. Expression and purification of chicken beta interferon and its antivirus immunological activity[J]. Protein Expr Purif,2012,84(1):123-129.
[9] 徐雷,冯若飞,李向茸,等.猪IFN-β真核表达载体pEGFP-C1-IFN-β的构建及在CHO细胞中的表达[J]. 中国兽医科学,2015,45(7):751-757.
[10] LIN F C,YOUNG H A. Interferons:success in anti-viral immunotherapy[J]. Cytokine Growth Factor Rev,2014,25(4):369-376.
[11] HENSLEY L E,FRITZ L E,JAHRLING P B,et al. Interferon-beta 1a and SARS coronavirus replication [J]. Emerg Infect Dis,2004,10 (2):317-319.
[12] KOERNER I, KOCHS G, KALINKE U, et al. Protective role of beta interferon in host defense against influenza A virus[J]. J Virol,2007,81(4):2025-2030.
[13] IWATA A,SAITO T,MIZUKOSHI-LWATA N,et al. Cloning and expression of the canine interferon-beta gene[J]. J Interferon Cytokine Res,1996,16(10):765-770.
[14] 谭明芬. 犬IFN-β的基因克隆、原核表达及活性测定[D].扬州:扬州大学,2010.
[15] 张敏,张鑫宇,夏晓莉,等.类弹性蛋白多肽融合犬IFN-β的表达纯化和抗病毒活性及体外半衰期的研究[J].中国兽医科学,2016,46(12):1543-1549.