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| 猪圆环病毒Ⅰ型ORF1基因的克隆及其在大肠杆菌中的表达 | |
| 引用本文： | 马红艳,蒋智勇,孙丽华,郝永清,宋长绪. 猪圆环病毒Ⅰ型ORF1基因的克隆及其在大肠杆菌中的表达[J]. 中国兽医科学, 2007, 37(8): 666-669 |
| 作者姓名： | 马红艳 蒋智勇 孙丽华 郝永清 宋长绪 |
| 作者单位： | 1. 内蒙古农业大学,动物科学与医学学院,内蒙古,呼和浩特,010018;广东省农业科学院,兽医研究所,广东,广州,510640 2. 广东省农业科学院,兽医研究所,广东,广州,510640 3. 内蒙古农业大学,动物科学与医学学院,内蒙古,呼和浩特,010018 |
| 基金项目： | 国家食品安全重大专项项目;广东省农业科技攻关项目;广东省自然科学基金;广东省自然科学基金 |
| 摘 要： | 根据GenBank中猪圆环病毒Ⅰ型(PCV1)ORF1基因序列,设计合成了1对引物,对PCV1 ORF1基因进行PCR扩增,将扩增出的ORF1基因(939 bp)克隆入pMD18-T载体,将筛选获得的重组质粒命名为pMD-ORF1。测序分析表明,克隆的ORF1与广东分离株DQ659154的核苷酸序列同源性高达99.6%,推导的氨基酸序列同源性为99.4%。将ORF1双酶切产物插入原核表达载体pET-41a,得到的重组子命名为pET-ORF1。用IPTG进行诱导表达,收集菌液进行SDS-PAGE和Western-blot分析,结果表明,PCV1ORF1在pET-41a中获得了高效融合表达,其表达蛋白的分子质量约为67.6 ku。 |
| 关 键 词： | 克隆 表达 |
| 文章编号： | 1673-4696(2007)08-0666-04 |
| 修稿时间： | 2007-01-26 |
Cloning of ORF1 gene of porcine circovirus type 1 and its expression in Escherichia coli |
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| MA Hong-yan,JIANG Zhi-yong,SUN Li-hua,HAO Yong-qing,SONG Chang-xu. Cloning of ORF1 gene of porcine circovirus type 1 and its expression in Escherichia coli[J]. Chinese Veterinary Science, 2007, 37(8): 666-669 | |
| Authors: | MA Hong-yan JIANG Zhi-yong SUN Li-hua HAO Yong-qing SONG Chang-xu |
| Abstract: | A pair of primers was designed according to the porcine circovirus type 1(PCV1) sequences in GenBank to amplify the open reading frame 1(ORF1) gene of PCV1.Then,the amplicon was cloned into pMD18-T vector to construct recombinant plasmid pMD-ORF1.The identity of nucleotide sequence was 99.6% between the amplified ORF1 gene and that of Guangdong isolate DQ659154,and the identity of the deduced amino acid sequence was 99.4% correspondingly.The ORF1 gene was subcloned into a prokaryo-tic expression vector pET-41a and the recombinant was designated pET-ORF1.The results of SDS-PAGE and Western-blotting indicated that the ORF1 gene was expressed from pET-ORF1 induced by IPTG at high level,and the expressed fusion protein was 67.6 ku approximately in molecular mass. |
| Keywords: | PCV1ORF1 |
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