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| 猪圆环病毒2型ORF1和ORF2基因的克隆与表达 | |
| 引用本文： | 王生育,颜江华,孔繁德,姜海军,张长弓,黄印尧.猪圆环病毒2型ORF1和ORF2基因的克隆与表达[J].中国预防兽医学报,2007,29(12):934-937. |
| 作者姓名： | 王生育 颜江华 孔繁德 姜海军 张长弓 黄印尧 |
| 作者单位： | 1. 厦门大学,医学院抗癌研究中心,福建,厦门,361005 2. 厦门出入境检验检疫局,福建,厦门,361012 3. 福建农林大学,动物科学学院,福建,福州,361006 |
| 摘 要： | 根据GenBank猪圆环病毒2型基因序列,设计合成两对特异性引物,对厦门株-1进行PCR扩增,得到ORF1和ORF2基因。将PCR产物酶切后,插入到pET-22b载体中,构建原核表达载体pET-22b/ORF1、pET-22b/ORF2,转化大肠杆菌BL21(DE_3),经IPTG诱导表达,收集茵液进行SDS-PAGE分析。确定重组蛋白主要以包涵体的形式表达,包涵体洗涤溶解后,采用Ni~(2 )离子金属螯合亲和层析柱纯化蛋白,逐步透析法进行复性。Western blot和ELISA分析结果表明Rep蛋白和Cap蛋白均能与猪圆环病毒2型阳性血清发生特异性反应。 |
| 关 键 词： | 猪圆环病毒 Rep蛋白 Cap蛋白 表达 |
| 文章编号： | 1008-0589（2007）12-0934-04 |
| 收稿时间： | 2007-02-18 |
| 修稿时间： | 2007年2月18日 |
Cloning and expression of ORF1 and ORF2 genes of porcine circovirus type 2 |
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| WANG Sheng-yu,YAN Jiang-hua,KONG Fan-de,JIANG Hai-jun,ZHANG Chang-gong,HUANG Yin-yao.Cloning and expression of ORF1 and ORF2 genes of porcine circovirus type 2[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(12):934-937. | |
| Authors: | WANG Sheng-yu YAN Jiang-hua KONG Fan-de JIANG Hai-jun ZHANG Chang-gong HUANG Yin-yao |
| Abstract: | ORF1 and ORF2 genes of porcine circovirus type 2(PCV-2)were obtained by PCR with specific primers designed according to PCV-2 sequence in GenBank and cloned into pET-22b,respectively.The resultant constructs were transformed into E.coli BL21(DE3)and expressed by IPTG induction.The expression products,which existed mainly as inclusion bodies,was purified by Ni~(2 )-affinity chromatographic columnwas and refolded following dialysis.Western blot and ELISA analysis showed that the purified products could react with PCV-2 specific serum. |
| Keywords: | porcine circovirus Rep protein Cap protein expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |