Binding of Grass Carp(Ctenopharyngodon idellus) Invariant Chain(Ii)with Its Major Histocompatibility Complex(MHC) Molecules Detected with a Pull-down Method
恒定链(invariant chain,Ii)能结合主要组织相容性复合体(major histocompatibility complex,MHC)Ⅰ类和Ⅱ类分子,并在MHC递呈抗原中起重要辅助作用。为了探讨鱼类Ii和MHC分子体外结合关系,本研究采用Pull-down方法检测草鱼(Ctenopharyngodon idellus)Ii与MHCⅠ类或Ⅱ类分子的结合。克隆草鱼Ii(711 bp)、MhcⅠα(540 bp)和MhcⅡβ(549 bp)基因片段,再将其分别插入原核表达质粒PET-22b(无His标签)或PET-28a(含His标签)中,构建了3个重组质粒PET-22b-Ii、PET-28a-MhcⅠα和PET-28aMhcⅡβ。然后分别转染工程菌大肠杆菌(Escherichia coli)Rosetta(DE3),进行诱导表达、纯化和聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,PAGE)鉴定。结果表明,含His标签的MHCⅠα和MHCⅡβ不仅表达,而且可高效纯化,其相对分子质量约为24 k D;Western blot结果表明,Ii可表达,相对分子质量...
【英文摘要】
Invariant chain(Ii) can bind to major histocompatibility complex(MHC) classⅠorⅡmolecules,and plays an important assistant role in antigen presentation by MHC molecules. In order to explore what about the binding of Ii with MHC molecules, a Pull- down method for detection of association of grass carp(Ctenopharyngodon idellus) Ii with MHC molecules was reported in the present study. DNA segments of Ii(711 bp), MhcⅠα(540 bp) and MhcⅡβ(549 bp) in grass carp were cloned, and then respectively inserted intoprokar...
【基金】
国家自然科学基金(No.31201888和No.31172306)
【更新日期】
2015-07-08
【分类号】
Q78
【正文快照】
主要组织相容性复合体(major histocompatibil-ity complex,MHC)Ⅰ类和Ⅱ类分子分别递呈内源性和外源性抗原肽,在启动特异性免疫中起关键作用。在Ⅱ类分子递呈外源性抗原肽中,恒定链(in-variant chain,Ii)是重要的伴侣分子(Jasanoff et al.,1998),参与MHCⅡ类分子成熟和装配(St
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