鸡MHCⅠ分子二聚体融合基因的构建及表达

doi:10.11924/j.issn.1000-6850.casb16010120

中国农学通报 ›› 2016, Vol. 32 ›› Issue (20): 1-5.doi: 10.11924/j.issn.1000-6850.casb16010120

所属专题：生物技术；畜牧兽医

• 畜牧动物医学蚕蜂 • 下一篇

鸡MHCⅠ分子二聚体融合基因的构建及表达

戴银,胡晓苗,赵瑞宏,侯宏艳,沈学怀,潘孝成,周学利,张丹俊

(安徽省农业科学院畜牧兽医研究所,合肥 230031）

收稿日期:2016-01-24 修回日期:2016-03-18 接受日期:2016-03-25 出版日期:2016-07-15 发布日期:2016-07-15
通讯作者: 张丹俊
基金资助:
国家自然科学基金项目“MHCⅠ分子靶向性疫苗载体的构建及其免疫作用的机理”(31302044)；国家现代农业产业技术体系专项“合肥综合试验站”(CARS-41)；安徽省科技攻关项目“鸡J-亚群白血病综合防治技术研究”(11010302119)。

Construction and Expression of Fusion Gene of Chicken MHCⅠ Dimers

Dai Yin, Hu Xiaomiao, Zhao Ruihong, Hou Hongyan, Shen Xuehuai, Pan Xiaocheng, Zhou Xueli, Zhang Danjun

(Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Science, Hefei 230031)

Received:2016-01-24 Revised:2016-03-18 Accepted:2016-03-25 Online:2016-07-15 Published:2016-07-15

摘要/Abstract

摘要： 为进一步研究主要组织相容性复合体Ⅰ类分子(major histocompatibility complex, MHCⅠ)的作用机制，提高其抗原呈递的效率，获得生物活性较好的鸡MHCⅠ分子二聚体，采用4个不同长度的连接肽G₄S、(G₄S)₂、(G₄S)₃和(G₄S)₄，通过基因重组技术将MHCⅠα和β₂m链基因共价连接，随后插入带有绿色荧光标签蛋白的真核表达载体，成功构建了4个重组质粒pEGFP-MHCⅠβ₂m-α。真核细胞表达后，荧光显微镜观察可见，各重组MHCⅠ二聚体分子均主要分布于真核细胞的内膜系统。此外，Western Blot检测均可见蛋白大小约80.0 kD左右的特异反应带，表达蛋白与鸡MHCⅠ类分子的相应抗体可发生特异性结合反应。研究结果表明各连接肽的重组质粒pEGFP-MHCⅠβ₂m-α均能够在真核细胞中顺利表达，且融合蛋白具有良好的免疫学活性。

关键词: 翻耕深度, 翻耕深度, 农艺性状, 产量, 棉花, 新疆

Abstract: To explore the mechanism of major histocompatibility complex classⅠ (MHCⅠ), improve the efficiency of antigen presentation, the dimers of chicken MHC molecules were obtained. The MHCⅠ α and β₂m chain gene were linked by different length linkers with sequences encoding G₄S, (G₄S)₂, (G₄S)₃and (G₄S)₄. Then this fragments were inserted into the eukaryotic expression vector with green fluorescent protein, and the recombinant plasmids (pEGFP-MHCⅠβ₂m-α) were constructed. The recombinant plasmids were transfected into eukaryotic cells. The gene products of recombinant plasmids were mainly located at endomembrane system of the eukaryotic cells under fluorescence microscopy. Moreover, the positive reactions were observed by Western Blot, and all the proteins had a molecular weight of about 80.0 kD, in accordance with the target proteins, and the proteins had binding reaction with specific antibodies. The results showed that the recombinant plasmids with different length linkers could be expressed in eukaryotic cells with a good immunological activity.

戴银,胡晓苗,赵瑞宏,侯宏艳,沈学怀,潘孝成,周学利,张丹俊. 鸡MHCⅠ分子二聚体融合基因的构建及表达[J]. 中国农学通报, 2016, 32(20): 1-5.

Dai Yin,Hu Xiaomiao,Zhao Ruihong,Hou Hongyan,Shen Xuehuai,Pan Xiaocheng,Zhou Xueli and Zhang Danjun. Construction and Expression of Fusion Gene of Chicken MHCⅠ Dimers[J]. Chinese Agricultural Science Bulletin, 2016, 32(20): 1-5.

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鸡MHCⅠ分子二聚体融合基因的构建及表达

Construction and Expression of Fusion Gene of Chicken MHCⅠ Dimers

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