Construction of Eukarytic Expression Vector Containing PCV-2 ORF2 and Its Expression

TAN Guo—lei; HU Zhi—hua; LU Ying—hua; CHEN De—sheng; CHEN Pu—yan

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August 2003

TAN Guo—lei, HU Zhi—hua, LU Ying—hua, CHEN De—sheng and CHEN Pu—yan. Construction of Eukarytic Expression Vector Containing PCV-2 ORF2 and Its Expression[J]. Virologica Sinica, 2003, 18(4): 367-370.

Citation: 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 .VIROLOGICA SINICA, 2003, 18(4) : 367-370.

猪圆环病毒2型ORF2编码基因真核表达载体的构建及表达

1.
南京农业大学动物医学院，江苏南京，210095

摘要

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plREShyg载体上，构建了plRESiORF2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plRESiORF2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。
- 猪圆环病毒
- , ORF2：信号肽

Construction of Eukarytic Expression Vector Containing PCV-2 ORF2 and Its Expression

1.
College ofVeterinary Medicine，Nanjing Agriculture Universityt Nanfing 210095，China

Abstract

According to the published OI 2 gene sequence of PCV一2．two primers were designed to amplify the ORF2 gene from PK一1 5 cells inoculated by PCV一2．It was cloned into the pSecTag2 vector,and the recombinant plasmid named pSecORF2 was obtained．Another primer was designed to amplify OI 2 gene including signal peptide sequence from pSecORF2．Then it was cloned into pIREShyg vector．Th e recombinan t expression vector plRESiORF2 was constructed．CHO cells were tran sfected in vitro with them． Th e expression of ORF2 gene was detected by indirect immunofluorescence assay(If1A、．Further research iS underway to study the biological activity of OI protein an d to establish the diagnostic kit of PCV
- Porcine circovirus
- , ORF2：Signal peptide

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction of Eukarytic Expression Vector Containing PCV-2 ORF2 and Its Expression

1. College ofVeterinary Medicine，Nanjing Agriculture Universityt Nanfing 210095，China

Abstract: According to the published OI 2 gene sequence of PCV一2．two primers were designed to amplify the ORF2 gene from PK一1 5 cells inoculated by PCV一2．It was cloned into the pSecTag2 vector,and the recombinant plasmid named pSecORF2 was obtained．Another primer was designed to amplify OI 2 gene including signal peptide sequence from pSecORF2．Then it was cloned into pIREShyg vector．Th e recombinan t expression vector plRESiORF2 was constructed．CHO cells were tran sfected in vitro with them． Th e expression of ORF2 gene was detected by indirect immunofluorescence assay(If1A、．Further research iS underway to study the biological activity of OI protein an d to establish the diagnostic kit of PCV