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编码Fc受体的小鼠巨细胞病毒早期基因的鉴定与表达

Identification and expression of a murine cytomegalovirus early gene coding for an Fc receptor.

作者信息

Thäle R, Lucin P, Schneider K, Eggers M, Koszinowski U H

机构信息

Department of Virology, University of Heidelberg, Germany.

出版信息

J Virol. 1994 Dec;68(12):7757-65. doi: 10.1128/JVI.68.12.7757-7765.1994.

Abstract

Several herpesviruses, including cytomegalovirus, induce receptors for the Fc domain of murine immunoglobulin G (IgG) molecules. Viral genes coding for these receptors have been characterized only for alphaherpesviruses. In this report, we describe a new approach that led to the identification of an Fc receptor (FcR) of murine cytomegalovirus (MCMV). The Fc fragment of IgG precipitated glycoproteins (gp) of 86 to 88 and 105 kDa from MCMV-infected cells. Deglycosylation by endoglycosidase F resulted in a protein with a molecular mass of 64 kDa. Injection of complete MCMV DNA or of DNA fragments, and the subsequent testing of cytoplasmic binding of IgG by immunofluorescence microscopy, was used to search for the coding region in the MCMV genome. The gene was located in the HindIII J fragment, map units 0.838 to 0.846, where an open reading frame of 1,707 nucleotides predicts a gp of 569 amino acids with a calculated molecular mass of 65 kDa. The sequence of this gp is related to those of the gE proteins of herpes simplex virus type 1 and varicella-zoster virus. The defined length of the mRNA, 1,838 nucleotides, was in agreement with that of a 1.9-kb RNA expressed throughout the replication cycle, starting at the early stages of infection. Expression of the gene fcr1 by recombinant vaccinia virus resulted in the synthesis of gp86/88 and gp105, each with FcR properties, and the correct identification of the gene encoding the FcR was confirmed by the DNA injection method.

摘要

包括巨细胞病毒在内的几种疱疹病毒可诱导小鼠免疫球蛋白G(IgG)分子Fc结构域的受体。编码这些受体的病毒基因仅在α疱疹病毒中得到了表征。在本报告中,我们描述了一种新方法,该方法导致鉴定出小鼠巨细胞病毒(MCMV)的一种Fc受体(FcR)。IgG的Fc片段从感染MCMV的细胞中沉淀出86至88 kDa和105 kDa的糖蛋白(gp)。内切糖苷酶F去糖基化后产生一种分子量为64 kDa的蛋白质。注射完整的MCMV DNA或DNA片段,随后通过免疫荧光显微镜检测IgG的细胞质结合情况,以此在MCMV基因组中寻找编码区域。该基因位于HindIII J片段,图谱单位为0.838至0.846,其中一个1707个核苷酸的开放阅读框预测编码一个569个氨基酸的gp,计算分子量为65 kDa。该gp的序列与单纯疱疹病毒1型和水痘带状疱疹病毒的gE蛋白序列相关。确定的mRNA长度为1838个核苷酸,与在整个复制周期中表达的1.9 kb RNA的长度一致,该RNA从感染早期开始表达。重组痘苗病毒表达基因fcr1导致合成具有FcR特性的gp86/88和gp105,并且通过DNA注射法证实了对编码FcR的基因的正确鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6f9/237237/cb838a01e83c/jvirol00021-0105-a.jpg

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