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当前位置: 首页 > 文章 > 表达禽流感病毒NP和M2融合基因重组乳酸菌的构建及鉴定 中国兽药杂志 2018 (12) 9-15
Position: Home > Articles > Construction and Identification of Recombinant Lactobacillus plantarum NC8 Expressing NP Fused with M2 Proteins of Avian Influenza Virus Chinese Journal of Veterinary Drug 2018 (12) 9-15

表达禽流感病毒NP和M2融合基因重组乳酸菌的构建及鉴定

作  者：
李琼燕;杨桂连;张成赛;赵金辉;晋玉北;杨文涛;王春凤
单  位：
吉林农业大学动物科学技术学院吉林省动物微生态制剂工程研究中心
关键词：
禽流感病毒;NP基因;M2基因;原核表达;重组乳酸菌
摘  要：
为了在乳酸菌中表达H9N2亚型禽流感病毒(AIV) NP和M2基因并检测其反应原性,先以pMD19-T-M2质粒作为模板,采用PCR方法克隆M2全长基因,然后将M2基因亚克隆入p SIP409-pgsA'-NP质粒中,构建重组质粒pSIP409-pgsA'-NP-M2,再将重组质粒pSIP409-pgsA'-NP-M2通过电转化的方法转入植物乳杆菌Lb.plantarum NC8中,制备重组菌NC8-p SIP409-pgsA'-NP-M2并诱导表达,最后通过流式细胞术、免疫荧光和免疫印迹技术验证。结果表明,重组菌NC8-pSIP409-pgsA'-NP-M2能够经诱导表达NP-M2融合蛋白,并且与兔源NP单克隆抗体和抗M2的小鼠血清具有反应原性。试验为后续研究禽流感广谱口服疫苗奠定了基础。
译  名：
Construction and Identification of Recombinant Lactobacillus plantarum NC8 Expressing NP Fused with M2 Proteins of Avian Influenza Virus
作  者：
LI Qiong-yan;YANG Gui-lian;ZHANG Cheng-sai;ZHAO Jin-hui;JIN Yu-bei;YANG Wen-tao;WANG Chun-feng;College of Animal Science and Technology,Jilin Agricultural University,Jilin Provincial Engineering Research Center of Animal Probiotics;
单  位：
LI Qiong-yan%YANG Gui-lian%ZHANG Cheng-sai%ZHAO Jin-hui%JIN Yu-bei%YANG Wen-tao%WANG Chun-feng%College of Animal Science and Technology,Jilin Agricultural University,Jilin Provincial Engineering Research Center of Animal Probiotics
关键词：
avian influenza virus;;NP gene;;M2 gene;;prokaryotic expression;;recombinant bacteria
摘  要：
To express the M2-NP fusion gene of the H9 N2 subtype avian influenza virus and to study the reactivity of its prokaryotic expression product in Lb. plantarum NC8,p MD19-T-M2 plasmid supplied from the laboratory serves as the template,the M2 gene was then amplified by PCR. The M2 gene was subcloned in the pSIP409-pgsA '-NP vector to construct a recombinant plasmid pSIP409-pgsA '-NP-M2. The recombinant plasmid pSIP409-pgsA '-NP-M2 was transformed into Lb. plantarum NC8 by electroporation to prepare recombinant bacteria NC8-p SIP409-pgsA'-NP-M2. The NC8-pSIP409-pgsA'-NP-M2 was induced,and reactogenicity of antigens were detected using flow cytometry,immunofluorescence and Western-blot. The results showed that NP-M2 fusion protein was able to express on NC8-p SIP409-pgsA'-NP-M2. Antigens of recombinant bacteria were also recognized with anti-NP monoclonal antibody and anti-M2 mouse source serum,which play an important role on developing universal oral vaccine against avian influenza viruses.

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