| 猪圆环病毒2型Cap蛋白在杆状病毒系统表达及鉴定 |
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| 引用本文： | 宋月,姜艳萍,崔文,李一经. 猪圆环病毒2型Cap蛋白在杆状病毒系统表达及鉴定[J]. 东北农业大学学报, 2012, 43(3): 36-41 |
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| 作者姓名： | 宋月 姜艳萍 崔文 李一经 |
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| 作者单位： | 东北农业大学动物医学学院,哈尔滨,150030 |
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| 基金项目： | 国家十一五支撑项目(2009BADB4B01) |
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| 摘 要： | 通过PCR方法扩增猪圆环病毒2型(pcv-2)中的缺失核定位序列的ORF2基因,将其克隆到Bac-N-Blue杆状病毒表达系统的pMelBacB载体中,得到阳性重组质粒pMel-ORF2。将其与Bac-N-Blue DNA体外重组,在脂质体的介导下,转染处于对数生长期的Sf9细胞,获得重组杆状病毒并在Sf9细胞上表达重组蛋白。通过SDS-PAGE、Western-blot和间接免疫荧光法(IFA)鉴定表明,Cap蛋白在昆虫-杆状病毒系统获得了表达,为Cap蛋白作为pcv-2检测抗原进行猪圆环病毒特异性抗体的检测提供了物质基础。
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| 关 键 词： | 猪圆环病毒2型Cap蛋白 重组杆状病毒 Sf9细胞 |
Expression and identification of capsid protein of porcine circovirus type 2 by recombinant baculovirus system |
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| SONG Yue , JIANG Yanping , CUI Wen , LI Yijing. Expression and identification of capsid protein of porcine circovirus type 2 by recombinant baculovirus system[J]. Journal of Northeast Agricultural University, 2012, 43(3): 36-41 |
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| Authors: | SONG Yue JIANG Yanping CUI Wen LI Yijing |
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| Affiliation: | (College of Veterinary Medicines,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | The ORF2 gene of Porcine Circovirus type 2(PCV2) deleting N-terminal nuclear localization region was amplified by PCR and cloned into the donor vector pMelBacB of Bac-N-Blue Baculovirus Expression System,and the recombinant plasmid was verified.After homologous recombination of the plasmid with Bac-N-Blue linear DNA,the mixture was transfected into Sf9 insect cells by Cellfectin Reagent,and the recombinant Baculovirus was constructed,which expressed recombinant protein on Sf9 cell.The result of confirming by SDS-PAGE,Western-blot and indirect lmmunofluorescence,indicated the recombinant Cap protein was expressed by Baculovirus system.It provided a substance foundation to test the specific antibody of Porcine Circovirus with Cap protein as detection antigen. |
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| Keywords: | capsid protein of porcine circovirus type 2 recombinant baculovirus Sf9 cell |
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