| 摘要: |
| 利用RT-PCR技术扩增出不含信号肽的猪瘟病毒石门株的E2基因,将其克隆到PGEX-T-Easy 载体上,用BamHⅠ和Hind Ⅲ进行双酶切,回收目的基因。将目的基因克隆到表达载体质粒pPROEX-HTb中,获得重组质粒pPROEXE2,用pPROEXE2转化大肠杆菌,诱导含重组质粒pPROEXE2的大肠杆菌BL21表达E2基因蛋白,研究E2蛋白与猪瘟阳性血清反应的特异性。结果表明,受体菌诱导后能表达E2基因蛋白,所表达的E2蛋白能与猪瘟阳性血清发生特异性很强的反应。 |
| 关键词: 猪瘟病毒石门株 E2基因 克隆 表达 |
| DOI： |
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| 基金项目:陕西省科技攻关项目(2003K02-G11) |
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| Study on the cloning and expression of E2 gene of Classical swine fever virus in E.coli |
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| Abstract: |
| The E2 gene of CSFV Shimen strain was amplified by reverse transcription polymerase chain reaction (RT-PCR). The amplified fragments were cloned into PGEX-T-Easy vector. We had digested the interested gene by the property without both of the two cleavage sites of PGEX-T-Easy vector and obtained the recombinant plasmids pPROEXE2 by cloning the interested gene into the expressed vector plas-mids pPROEX-HTb. It was identified that the position of the E2 gene insert,the size and the reading frame were all right by PCR, restriction digestion and the sequence analysis. SDS-PAGE indicated that the recipient germs induced by the recombinant plasmids could express E2 gene of CSFV. Western blot indicated that the positive serum of CSFV could recognize the expressed antigen protein. |
| Key words: classical swine fever virus shimen strain E2 gene cloning expression |
