| H5N1亚型禽流感病毒血凝素蛋白单链抗体基因的克隆及表达 |
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| 引用本文： | 郝淑美,李群,王承宇,王化磊,杨松涛,夏咸柱.H5N1亚型禽流感病毒血凝素蛋白单链抗体基因的克隆及表达[J].病毒学报,2009,25(1):63-67. |
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| 作者姓名： | 郝淑美 李群 王承宇 王化磊 杨松涛 夏咸柱 |
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| 作者单位： | 长春理工大学,生命科学技术学院,长春,130022;解放军军事医学科学院,军事兽医研究所,长春,130062;解放军军事医学科学院,军事兽医研究所,长春,130062;解放军军事医学科学院,军事兽医研究所,长春,130062;吉林大学,畜牧兽医学院,长春,130062 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)课题(2005CB523005) |
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| 摘 要： | 构建并表达H5N1亚型禽流感病毒血凝素蛋白单链抗体,为禽流感靶向治疗药物的研制制备靶向载体。从分泌血凝素单克隆抗体的杂交瘤细胞株中提取mRNA,采用RT-PCR法扩增出重链和轻链可变区基因,通过SOE-PCR法将重链和轻链通过Linker连接起来构建单链抗体基因,将获得的单链抗体基因装入原核表达载体pET28a(+)中,构建重组质粒并表达,以Western blot鉴定单链抗体的特异性。结果成功构建了单链抗体基因,全长714bp,经原核表达,所构建的单链抗体可与H5亚型禽流感病毒HA蛋白特异结合,为禽流感的靶向治疗奠定了基础。
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| 关 键 词： | 禽流感病毒 血凝素蛋白 单链抗体 克隆 表达 |
Cloning and Expression of Single-Chain Fv Antibodies Against H5N1 Avian Influenza Virus Hemagglutinin |
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| HAO Shu-mei,LI Qun,WANG Cheng-yu,WANG Hua-lei,YANG Song-tao,XIA Xian-zhu.Cloning and Expression of Single-Chain Fv Antibodies Against H5N1 Avian Influenza Virus Hemagglutinin[J].Chinese Journal of Virology,2009,25(1):63-67. |
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| Authors: | HAO Shu-mei LI Qun WANG Cheng-yu WANG Hua-lei YANG Song-tao XIA Xian-zhu |
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| Institution: | 1.Changchun University of Science and Technology;Changchun 130022;China;2.Institute of Military Veterinary;Academy of Military Medical Sciences of PLA;Changchun 130062;3.College of Animal Science and Veterinary Medicin;Jilin University;changchun 130062;China |
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| Abstract: | To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension(SOE) with(Gly4Ser)3 linker.The recombinant plasmid was then transformed to E.coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714bp and the expression of Fv was validated by PAGE and Western blot. |
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| Keywords: | AIV hemagglutinin scFv cloning expression |
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