| 猪圆环病毒2型Cap羧基端多肽原核表达及抗原性分析 |
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| 引用本文： | 俞莉俐,姜平,冯志新.猪圆环病毒2型Cap羧基端多肽原核表达及抗原性分析[J].畜牧与兽医,2006,38(11):14-17. |
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| 作者姓名： | 俞莉俐 姜平 冯志新 |
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| 作者单位： | 南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏,南京,210095 |
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| 摘 要： | 本研究用PCR方法获得猪圆环病毒2型(PCV2)Cap蛋白基因,经酶切后将ORF2 3′端部分基因克隆至原核表达载体pGEX-6p-1中,转化大肠杆菌BL21,加入终浓度为0.3 mmol/LIPTG,在30℃诱导表达6 h后,经SDS-PAGE及western-blot分析,目的基因得到正确表达,大小约为40 ku。收集菌体并进行超声裂解后证实,表达产物以可溶蛋白与包涵体共同存在。对可溶蛋白与包涵体分别进行纯化,分别用做ELISA包被抗原检测PCV2抗体,结果两者无明显差异。
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| 关 键 词： | 猪圆环病毒2型 重组Cap蛋白 包涵体 ELISA |
| 文章编号： | 0529-5130（2006）11-0014-04 |
| 收稿时间： | 03 13 2006 12:00AM |
| 修稿时间： | 2006-03-13 |
Prokaryotic expression of C-terminal peptide of PCV2 Cap protein and immunogenicity analysis |
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| YU Li-li,JIANG Ping,FENG Zhi-xin.Prokaryotic expression of C-terminal peptide of PCV2 Cap protein and immunogenicity analysis[J].Animal Husbandry & Veterinary Medicine,2006,38(11):14-17. |
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| Authors: | YU Li-li JIANG Ping FENG Zhi-xin |
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| Institution: | Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agricuhure, Nanjing Agricultural University, Nanjing 210095, China |
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| Abstract: | ORF2 gene of porcine circovirus type 2(PCV2) was amplified by PCR.The recombinant expression vector pGEX-V2 was constructed by cloning parts of C-terminal of the PCR product into prokaryotic expression plasmid pGEX-6p-1.The recombinant vector was transformed into E.coli BL21 strain.With the IPTG induction concentration of 0.3 mmol/L,the fusion protein was analyzed with SDS-PAGE and Western-blotting 6 hours after induction at 30 ℃.The results revealed that the product with about 40 ku existing as inclusion-body and soluble protein had been expressed correctly.They were compared as coating antigens in ELISA detection after purified.There were no apparently differences in detecting PCV2 antibodies. |
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| Keywords: | porcine circovirus type 2 recombinant Cap protein inclusion-body ELISA |
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