山羊痘病毒A6L基因的克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2015.08.008

›› 2015, Vol. 42 ›› Issue (8): 1973-1978.doi: 10.16431/j.cnki.1671-7236.2015.08.008

山羊痘病毒A6L基因的克隆及序列分析

赵银龙, 吴国华, 朱海霞, 吴健, 颜新敏, 赵志荀, 李健, 吴娜, 穆晓峰, 张强

中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 农业部畜禽病毒学重点开放实室验, 兰州 730046

收稿日期:2015-02-02 出版日期:2015-08-20 发布日期:2015-08-27
通讯作者: 吴国华 E-mail:wuguohua@caas.cn
作者简介:赵银龙(1989-),男,河北邢台人,硕士生,研究方向:动物营养与微生物营养,E-mail:515911538@qq.com
基金资助:
国家质检总局公益性行业科研专项(201310093);国家自然科学基金(31201892);甘肃省自然科学基金(1208RJZA101);国家863计划(2012AA101304)

Cloning and Sequence Analysis of A6L Gene of Goatpox Virus

ZHAO Yin-long, WU Guo-hua, ZHU Hai-xia, WU Jian, YAN Xin-min, ZHAO Zhi-xun, LI Jian, WU Na, MU Xiao-feng, ZHANG Qiang

Key Laboratory of Animal Virology of the Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2015-02-02 Online:2015-08-20 Published:2015-08-27

摘要/Abstract

摘要： 为了分析山羊痘病毒A6蛋白的分子特征,本试验提取了山羊痘病毒古浪分离株的基因组DNA,设计引物进行PCR扩增,克隆A6L基因的DNA序列。将PCR产物连接到pGEM-T Easy载体后转化至大肠杆菌DH5α感受态细胞,筛选阳性克隆进行序列测定,利用生物信息学软件对A6L基因序列进行预测分析。结果显示,A6L基因序列含有1个由1128个核苷酸组成的开放阅读框,编码375个氨基酸残基组成的多肽,蛋白质分子质量的理论值为43.68 ku,理论等电点为7.50。该蛋白的二级结构中,α螺旋占53.30%,β折叠占10.24%,其余39.46%为无规则卷曲。多序列比对分析结果显示,不同羊痘病毒分离株A6L序列高度保守,同源性在97%以上。上述研究结果为进一步研究A6蛋白的生物学功能和羊痘病毒早期蛋白的分子相互作用奠定了基础。

关键词: 山羊痘病毒; A6L基因; 基因克隆; 序列分析

Abstract: To explore the molecular characteristics of protein A6 from goatpox virus (GTPV), genomic DNA was extracted from goatpox virus Gulang isolate.The specific primers were designed and used to amplify the A6L gene from the genomic DNA by PCR.A PCR product was ligated into pGEM-T Easy vector.After transformation into E.coli DH5α, the positive clones were sequenced and the sequences were analyzed with the bioinformatics softwares.The result showed that A6L gene sequence contained an open reading frame (ORF) of 1128 nucleotides and deduced protein consisted of 375 amino acids with the theoretical molecular weight of 43.68 ku and isoelectric point of 7.50.Analysis of secondary structure of protein A6 revealed that α-helix, β-strand and random coil were 53.30%, 10.24% and 39.46%, respectively.Analysis of multiple sequence alignment indicated A6L gene from different capripoxvirus isolates were highly conserved with identity of more than 97%.The present results laid the foundation for further studies of biological functions of protein A6 and interaction among the early proteins of capripoxvirus.

Key words: goatpox virus; A6L gene; gene cloning; sequence analysis

中图分类号:

S852.65

赵银龙, 吴国华, 朱海霞, 吴健, 颜新敏, 赵志荀, 李健, 吴娜, 穆晓峰, 张强. 山羊痘病毒A6L基因的克隆及序列分析[J]. , 2015, 42(8): 1973-1978.

ZHAO Yin-long, WU Guo-hua, ZHU Hai-xia, WU Jian, YAN Xin-min, ZHAO Zhi-xun, LI Jian, WU Na, MU Xiao-feng, ZHANG Qiang. Cloning and Sequence Analysis of A6L Gene of Goatpox Virus[J]. , 2015, 42(8): 1973-1978.

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山羊痘病毒A6L基因的克隆及序列分析

Cloning and Sequence Analysis of A6L Gene of Goatpox Virus

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