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兔源四个编码ω干扰素基因的克隆与结构分析

Cloning and structural analysis of four genes encoding interferon-omega in rabbit.

作者信息

Charlier M, L'Haridon R, Boisnard M, Martal J, Gaye P

机构信息

Unité d'Endocrinologie de l'Embryon, I.N.R.A., Jouy-en-Josas, France.

出版信息

J Interferon Res. 1993 Oct;13(5):313-22. doi: 10.1089/jir.1993.13.313.

Abstract

By using an ovine interferon-tau (IFN-tau) cDNA probe, four recombinant phages were isolated from a rabbit genomic library and sequenced from nucleotides -450 to 1,300 relative to the CAP site. Each of the four rabbit genes contains an open reading frame of 595 nucleotides and code for proteins that exhibit structural characteristics of the interferon-omega (IFN-omega) family. They display more than 98% identity in their coding regions. The deduced amino acid sequences share > 96% sequence similarity. In contrast, the 5' and 3' noncoding regions have diverged considerably (approximately 50% identity). Amino acid comparisons of rabbit IFN-omega with IFN-omega of other species reveal the highest degree of identity with human (72%), followed by porcine (68%) IFN-omega. Rabbit IFN-omega displays only 57% sequence similarity with ovine IFN-tau. The coding regions of the four genes subcloned in a cytomegalovirus eukaryotic expression vector and transfected in monkey COS-7 cells direct the production of proteins that protect bovine and rabbit cells against vesicular stomatitis virus infection, thus demonstrating that these genes encode fully active IFN proteins. The expression of these genes was studied in Sendai-induced rabbit leukocytes. A single band of poly(A)+RNA hybridized with a rabbit IFN-omega probe under stringent conditions, whereas no IFN-omega transcript was detected with RNA isolated from uninduced leukocytes. Southern blot analysis suggest the existence of at least eight IFN-omega genes or pseudogenes in the rabbit genome.

摘要

通过使用绵羊干扰素 -τ(IFN -τ)cDNA 探针,从兔基因组文库中分离出四个重组噬菌体,并对相对于 CAP 位点的核苷酸 -450 至 1300 进行了测序。四个兔基因中的每一个都包含一个 595 个核苷酸的开放阅读框,并编码具有干扰素 -ω(IFN -ω)家族结构特征的蛋白质。它们在编码区域的同一性超过 98%。推导的氨基酸序列具有超过 96%的序列相似性。相比之下,5' 和 3' 非编码区域有很大差异(约 50%的同一性)。兔 IFN -ω 与其他物种的 IFN -ω 的氨基酸比较显示,与人类 IFN -ω 的同一性最高(72%),其次是猪 IFN -ω(68%)。兔 IFN -ω 与绵羊 IFN -τ 的序列相似性仅为 57%。将四个基因的编码区域亚克隆到巨细胞病毒真核表达载体中并转染到猴 COS -7 细胞中,可指导产生能保护牛和兔细胞免受水泡性口炎病毒感染的蛋白质,从而证明这些基因编码完全活性的 IFN 蛋白。在仙台病毒诱导的兔白细胞中研究了这些基因的表达。在严格条件下,一条单一的 poly(A)+RNA 带与兔 IFN -ω 探针杂交,而从未诱导的白细胞中分离的 RNA 未检测到 IFN -ω 转录本。Southern 印迹分析表明兔基因组中至少存在八个 IFN -ω 基因或假基因。

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