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牛疱疹病毒1型早期蛋白BICP27的鉴定与特性分析,该蛋白可能刺激mRNA 3'加工。

Identification and characterization of BICP27, an early protein of bovine herpesvirus 1 which may stimulate mRNA 3' processing.

作者信息

Singh M, Fraefel C, Bello L J, Lawrence W C, Schwyzer M

机构信息

Institute of Virology, Faculty of Veterinary Medicine, University of Zürich, Switzerland.

出版信息

J Gen Virol. 1996 Apr;77 ( Pt 4):615-25. doi: 10.1099/0022-1317-77-4-615.

Abstract

Sequence analysis of the left genomic terminus of bovine herpesvirus 1 (BHV-1) revealed two convergently transcribed genes with 3' ends about 300 bp apart. The gene on the left is the previously described circ gene; that on the right was found to encode a protein of 400 amino acids which was designated BICP27 because of its homology to ICP27 (Vmw63) of herpes simplex virus 1 (HSV-1) and related proteins from other alpha- beta- and gammaherpesviruses. Rabbit antisera raised against a synthetic oligopeptide representing the amino terminus of the predicted polypeptide demonstrated the presence of BICP27 in the nuclei of infected cells by in situ immunoadsorbent assays. In Western immunoblots, BICP27 was detected as a 50 kDa BHV-1 specific protein expressed with early kinetics, in contrast to HSV-1 ICP27 which is an immediate early (IE) protein. A DNA fragment containing BICP27 coding sequences was inserted into a baculovirus genome. The recombinant BICP27 protein, identified by its reactivity with the antipeptide sera, exhibited the same electrophoretic mobility as BICP27 specified by BHV-1. Transient expression assays using target genes differing only in their poly(A) sites showed that BICP27, like its HSV-1 counterpart, may be involved in 3' processing of mRNA.

摘要

牛疱疹病毒1型(BHV-1)基因组左末端的序列分析显示,有两个反向转录的基因,其3'末端相距约300 bp。左边的基因是先前描述的circ基因;右边的基因被发现编码一种由400个氨基酸组成的蛋白质,因其与单纯疱疹病毒1型(HSV-1)的ICP27(Vmw63)以及其他α、β和γ疱疹病毒的相关蛋白质具有同源性,故被命名为BICP27。通过原位免疫吸附试验,用针对代表预测多肽氨基末端的合成寡肽产生的兔抗血清证明了感染细胞核中存在BICP27。在Western免疫印迹中,与HSV-1的ICP27(一种立即早期(IE)蛋白)不同,BICP27被检测为一种以早期动力学表达的50 kDa BHV-1特异性蛋白。将包含BICP27编码序列的DNA片段插入杆状病毒基因组中。通过其与抗肽血清的反应性鉴定的重组BICP27蛋白,其电泳迁移率与BHV-1指定的BICP27相同。使用仅在其聚腺苷酸化位点不同的靶基因进行的瞬时表达试验表明,BICP27与其HSV-1对应物一样,可能参与mRNA的3'加工。

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