鸡MHCⅠα和β2m链基因真核表达载体的构建及其在293T细胞中的表达

doi:10.16431/j.cnki.1671-7236.2016.08.002

Abstract

Abstract:

To explore the mechanism of MHCⅠ molecule in immune response,chicken MHCⅠα and β₂m genes were cloned by PCR.Then the fragments were inserted into the eukaryotic expression vector with fluorescent protein,and the recombinant plasmids pEGFP-MHCⅠα and pmCherry-MHCⅠβ₂m were constructed.The recombinant plasmids were transfected into 293T cell with lipofectin reagent.The gene products of recombinant plasmids were mainly located to endomembrane system of the cells by fluorescence microscopy,and changed the intracellular localization of the fusion with the fluorescent protein.Moreover,the positive reactions were observed by the method of Western blotting,and the proteins had the molecular weight of 68.3 and 41.3 ku,respectively,in accord with the target proteins.The results showed that the recombinant plasmids were expressed in 293T cells with a good immunological activity,and the proteins had the binding reaction with specific antibodies.

Key words: chicken MHCⅠα gene; chicken MHCⅠβ₂m gene; eukaryotic expression vector; 293T cell

CLC Number:

Q786

DAI Yin, HU Xiao-miao, ZHAO Rui-hong, HOU Hong-yan, SHEN Xue-huai, PAN Xiao-cheng, ZHOU Xue-li, ZHANG Dan-jun. Construction of Eukaryotic Expression Vector of Chicken MHCⅠα and β₂m Genes and Its Expression in 293T Cells[J]. , 2016, 43(8): 1938-1943.

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Construction of Eukaryotic Expression Vector of Chicken MHCⅠα and β₂m Genes and Its Expression in 293T Cells

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Construction of Eukaryotic Expression Vector of Chicken MHCⅠα and β2m Genes and Its Expression in 293T Cells

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Construction of Eukaryotic Expression Vector of Chicken MHCⅠα and β₂m Genes and Its Expression in 293T Cells