| 猪圆环病毒2型ORF2编码基因真核表达载体的构建及表达 |
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| 引用本文： | 谈国蕾 胡志华 芦银华 陈德胜 陈溥言. 猪圆环病毒2型ORF2编码基因真核表达载体的构建及表达[J]. Virologica Sinica, 2003, 18(4): 367-370 |
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| 作者姓名： | 谈国蕾 胡志华 芦银华 陈德胜 陈溥言 |
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| 作者单位： | 南京农业大学动物医学院，南京农业大学动物医学院，南京农业大学动物医学院，南京农业大学动物医学院，南京农业大学动物医学院 江苏南京 210095，江苏南京 210095，江苏南京 210095，江苏南京 210095，江苏南京 210095 |
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| 基金项目： | 国家“863”项目资助(2001AA249012)，上海科技兴农重点攻关项目(农科攻字2001第3-5号) |
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| 摘 要： | 根据GenBank中猪圆环病毒2型(PCV-2)ORF2基因序列，设计了1对引物，应用PCR从含有PCV-2的PK-15细胞中扩增出ORF2基因，将其克隆入pSecTag2载体中，构建了pSecORF2载体。又设计一条含信号肽序列的上游引物，以pSecORF2为模板，扩增出含信号肽序列的ORF2基因，将其克隆到pIREShyg载体上，构建了pIRESiORF2真核表达载体。然后通过磷酸钙共沉淀法转染CHO细胞，进行表达。间接免疫荧光实验(IFA)成功检测到pIRESiORF2在CHO细胞中的表达。这为进一步研究ORF2编码蛋白的生物学活性及建立PCV诊断试剂盒打下基础。
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| 关 键 词： | 猪圆环病毒 ORF2 编码基因 真核表达载体 表达 |
Construction of Eukarytic Expression Vector Containing PCV-2 ORF2 and Its Expression |
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| TAN Guo-lei,HU Zhi-hua,LU Ying-hua,CHEN De-sheng,CHEN Pu-yan. Construction of Eukarytic Expression Vector Containing PCV-2 ORF2 and Its Expression[J]. 中国病毒学(英文版), 2003, 18(4): 367-370 |
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| Authors: | TAN Guo-lei HU Zhi-hua LU Ying-hua CHEN De-sheng CHEN Pu-yan |
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| Affiliation: | TAN Guo-lei,HU Zhi-hua,LU Ying-hua,CHEN De-sheng,CHEN Pu-yan~ |
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| Abstract: | According to the published ORF2 gene sequence of PCV-2, two primers were designed to amplify the ORF2 gene from PK-15 cells inoculated by PCV-2. It was cloned into the pSecTag2 vector, and the recombinant plasmid named pSecORF2 was obtained. Another primer was designed to amplify ORF2 gene including signal peptide sequence from pSecORF2. Then it was cloned into pIREShyg vector. The recombinant expression vector pIRESiORF2 was constructed. CHO cells were transfected in vitro with them. The expression of ORF2 gene was detected by indirect immunofluorescence assay(IFA). Further research is underway to study the biological activity of ORF2 protein and to establish the diagnostic kit of PCV. |
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| Keywords: | Porcine circovirus ORF2 Signal peptide |
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