| 鸡痘病毒ORF073基因克隆及原核表达 |
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| 引用本文： | 王彦红,马怀良,胡顺林,仇旭升,刘秀梵.鸡痘病毒ORF073基因克隆及原核表达[J].扬州大学学报(农业与生命科学版),2006,27(4):5-7,11. |
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| 作者姓名： | 王彦红 马怀良 胡顺林 仇旭升 刘秀梵 |
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| 作者单位： | 扬州大学,农业部畜禽传染病学重点开放实验室,江苏,扬州,225009 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 根据已经发表的鸡痘病毒(FPV)美国致病株的ORF073基因序列,设计1对引物,分别以282E4FPV、LP株FPV、102E6FPV为模板,扩增出目的片段,将其克隆到pGEM-TEasy载体中,经酶切鉴定证实,并进行序列测定。目的片段包含了大小为522 bp的ORF073基因,与GenBank上的AF198100(FPVUS)和AJ581527(FP9)序列进行序列比较分析表明:3种病毒株的ORF073基因与鸡痘病毒美国致病株(FPVUS)和英国弱毒株(FP9)完全一致。克隆的ORF073基因与原核表达载体pET-28a构建重组表达载体,经IPTG诱导,在BL21(DE3)细菌中表达了分子质量约32 ku的蛋白。
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| 关 键 词： | 鸡痘病毒 ORF073 基因克隆 原核表达 |
| 文章编号： | 1671-4652（2006）04-0005-03 |
| 收稿时间： | 2006-09-13 |
| 修稿时间： | 2006-09-13 |
Cloning and expression of ORF073 of fowlpox virus |
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| WANG Yan-hong,MA Huai-liang,HU Shun-ling,QIU Xu-shen,LIU Xiu-fan.Cloning and expression of ORF073 of fowlpox virus[J].Journal of Yangzhou University:Agricultural and Life Science Edition,2006,27(4):5-7,11. |
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| Authors: | WANG Yan-hong MA Huai-liang HU Shun-ling QIU Xu-shen LIU Xiu-fan |
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| Institution: | Key lab of Anita Inf Dis, MOA, Yangzhou Univ, Yangzhou, 225009, China |
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| Abstract: | DNA fragment(containing entire ORF073)amplified from fowlpox virus strain 282E4,102E6 and LP by PCR using primers based on DNA sequence of FPVUS,was cloned into PGEM-TEasy vector.The recombinant was indi- cated containing the complete open reading frame of ORF073 with 522 bp by sequencing.Nucleotide sequence comparisons found that ORF073 of 282E4,102E6 and LP was identical to FPVUS and FP9.The recombinant plasmid pET-28a- ORF073 was constructed and transformed into E.coli BL21(DE3).A recombinant protein with a relative molecular weight of 32 ku was expressed in E.coli. |
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| Keywords: | fowlpox virus ORF073 cloning expression |
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