Abstract：Bovine leukocyte antigen (BoLA), also named major histocompatibility complex (MHC), its class Ⅱ molecules and chaperone protein, invariant chain (Ii), plays a key role in the processing and presentation of antigenic peptides. In order to understand the characteristics of binding between BoLAⅡβ chain and Ii, and intracellular localization in this process, in this study, mRNA from blood cells of Huaibei yellow cattle (Bos taurus) was extracted and reverse transcribed into cDNA, the genes of BoLAⅡβ, its fragments and Ii were amplified by PCR and then the corresponding eukaryotic and prokaryotic expression plasmids were constructed, respectively. The proteins were expressed and purified by prokaryotic expression system. The binding between Ii and BoLAⅡβ chain or its fragments was detected by pull-down method and Western blot. The co-localization of Ii with BoLAⅡβ and their fragments in cells was observed by laser confocal microscope. The results showed that the recombinant plasmid containing Ii, BoLAⅡβ, BoLAⅡβ(β1β2) and BoLAⅡβ(TC) genes could be induced and purified. BoLAⅡβ and BoLAⅡβ(β1β2) had the active function of binding Ii, but BoLAⅡβ(TC) did not. Meanwhile, Ii could co-localized with BoLAⅡβ, BoLAⅡβ(β1β2) or BoLAⅡβ(TC) in eukaryotic cells. In conclusion, the β1β2 of BoLAⅡβ was the key domain for its intracellular binding and the both domains β1β2 and TC were the functional domains of co-localization with Ii in cells. This study provides an important experimental basis for further study on the transport and mechanism of BoLA molecule and Ii in immune response.
陈芳芳, 于凤梅, 张俊, 谈阳, 刘翠艳, 桂亚萍. 牛恒定链与BoLAⅡ类分子β链活性片段的结合与定位分析[J]. 农业生物技术学报, 2022, 30(10): 1884-1893.
CHEN Fang-Fang, YU Feng-Mei, ZHANG Jun, TAN Yang, LIU Cui-Yan, GUI Ya-Ping. Binding and Localization Analysis Between Bovine (Bos taurus) Invariant Chain and the Active Fragment of BoLA Class Ⅱ Molecule β Chain. 农业生物技术学报, 2022, 30(10): 1884-1893.
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