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Cloning and expression of the gene for an immunoglobulin G Fc receptor protein from a group A streptococcus.

机译:来自A组链球菌的免疫球蛋白G Fc受体蛋白的基因的克隆和表达。

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DNA containing the 5' end of the M-12 structural gene was used as a probe in colony hybridizations in an attempt to clone the M-76 gene from an M-type 76 group A streptococcal strain. A single positive colony was detected, and Southern hybridization analysis of plasmid DNA isolated from this colony indicated that the insert DNA had homology to the 5' end of the M-12 structural gene. Subclones were constructed to define the limits of the M-76 gene, and sonicates of these subclones were reacted with M-76-specific antiserum in immunodiffusion. A sonicate of one subclone, JM83(pDH56), reacted strongly with the M-76-specific antiserum but also reacted with preimmune rabbit serum. Protein expressed from this subclone bound immunoglobulin from horse and pig, as well as human myeloma immunoglobulin G (IgG) representing all four subclasses and purified human IgG Fc fragments. This indicated that JM83(pDH56) expressed a protein with characteristics previously attributed to the IgG Fc receptor protein from group A streptococci. Western blot analysis indicated that the cloned IgG Fc receptor protein had a molecular weight of approximately 29,000. Binding studies showed that the Fc receptor gene is expressed by the M-type 76 strain from which it was cloned and by an M- variant.
机译:试图从含有M-12结构基因5'端的DNA用作菌落杂交探针,以从M型76 A组链球菌菌株中克隆M-76基因。检测到单个阳性菌落,并且从该菌落分离的质粒DNA的Southern杂交分析表明,插入DNA与M-12结构基因的5'端具有同源性。构建亚克隆以定义M-76基因的限制,并在免疫扩散中将这些亚克隆的超声与M-76特异性抗血清反应。一个亚克隆,JM83(pDH56)的超声,与M-76特异性抗血清反应强烈,但也与免疫前的兔血清反应。从该亚克隆表达的蛋白质结合了来自人马的免疫球蛋白,以及代表所有四个亚类的人骨髓瘤免疫球蛋白G(IgG)和纯化的人IgG Fc片段。这表明JM83(pDH56)表达的蛋白具有先前归因于A组链球菌的IgG Fc受体蛋白的特征。 Western印迹分析表明,克隆的IgG Fc受体蛋白的分子量约为29,000。结合研究表明,Fc受体基因由克隆自其的M型76菌株和M变异体表达。

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