| 人巨细胞病毒pp150抗原区基因片段的克隆与表达 |
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| 引用本文： | 李国才,季明春,薛方明,严华,申厚凤.人巨细胞病毒pp150抗原区基因片段的克隆与表达[J].扬州大学学报(农业与生命科学版),2003,24(4):16-18. |
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| 作者姓名： | 李国才 季明春 薛方明 严华 申厚凤 |
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| 作者单位： | 1. 扬州大学,医学院,基础医学部,江苏,扬州,225001
2. 扬州大学,畜牧兽医学院,动物医学系,江苏,扬州,225009 |
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| 基金项目： | 江苏省卫生厅科研基金项目(9815);江苏省计划生育委员会科研基金项目;扬州大学科研基金项目(9911548) |
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| 摘 要： | 为了研制人巨细胞病毒(HCMV)核酸疫苗,根据HCMV基质磷蛋白pp150基因序列设计引物,利用高保真PCR从HCMV AD169株基因组DNA中扩增出编码pp150抗原决定簇区域的基因片段.序列分析结果显示其与发表的该段序列同源性为100%.将此基因片段克隆进原核表达质粒pET30a,重组质粒转化大肠杆菌BL21(DE3),诱导表达后经SDS-PAGE及Western blotting检测到约35 h的外源蛋白.表达产物免疫小鼠,所得抗体具有低滴度中和作用.
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| 关 键 词： | 人巨细胞病毒 pp150 抗原决定簇 原核表达 |
| 文章编号： | 1671-4652(2003)04-0016-03 |
| 修稿时间： | 2003年10月12 |
CLONING AND EXPRESSION OF THE GENE FRAGMENT ENCODING THE ANTIGENIC DETERMINANTS OF pp150 OF HUMAN CYTOMEGALOVIRUS |
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| LI Guo-cai,JI Ming-chun,XUE Fang-ming,YAN Hua,SHEN Hou-feng.CLONING AND EXPRESSION OF THE GENE FRAGMENT ENCODING THE ANTIGENIC DETERMINANTS OF pp150 OF HUMAN CYTOMEGALOVIRUS[J].Journal of Yangzhou University:Agricultural and Life Science Edition,2003,24(4):16-18. |
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| Authors: | LI Guo-cai JI Ming-chun XUE Fang-ming YAN Hua SHEN Hou-feng |
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| Abstract: | To develop a human cytomegalovirus (HCMV) DNA vaccine, the gene fragment encoding the antigenic determinants of pp150 was amplified from HCMV AD169 by PCR using a pair of primers based on the previously published gene sequence for HCMV pp150. The gene fragment was subcloned into pGEM-T vector and submitted to sequence analysis. DNA sequence alignment showed that it was 100% identical to that published. The gene fragment was then subcloned into a prokaryotic expression vector, pET30a, and a foreign protein about 35 ku was detected by SDS-PAGE and Western blotting assay after the recombinant plasmid was transformed into E.coli BL21 (DE3). The antiserum prepared by using the expressed product as antigen turned to be against pp150 specially by ELISA and had low neutralization effect. |
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| Keywords: | human cytomegalovirus pp150 antigenic determinant prokaryotic expression |
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