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人巨细胞病毒大型结构磷蛋白基因的图谱定位和核苷酸序列

Map position and nucleotide sequence of the gene for the large structural phosphoprotein of human cytomegalovirus.

作者信息

Jahn G, Kouzarides T, Mach M, Scholl B C, Plachter B, Traupe B, Preddie E, Satchwell S C, Fleckenstein B, Barrell B G

出版信息

J Virol. 1987 May;61(5):1358-67. doi: 10.1128/JVI.61.5.1358-1367.1987.

Abstract

Human cytomegalovirus particles contain a phosphoprotein of 150,000 (pp150) apparent molecular weight in their matrix; the protein appears particularly reactive in Western blot analyses with human antisera. The gene for pp150 was mapped by screening a bacteriophage lambda gt11 cDNA expression library with monospecific rabbit antisera. Subsequent hybridization of cDNA with cosmid and plasmid clones containing the human cytomegalovirus strain AD169 genome mapped the gene to HindIII fragments J and N. The gene is transcribed into a late 6.2-kilobase RNA. The nucleotide sequence of this region was determined, and a transcription initiation site and two polyadenylation sites of an abundant transcript were located by primer extension and nuclease protection experiments. The reading frame for pp150, deduced from computer analyses, gives rise to a polypeptide of 1,048 amino acids in length; protein secondary structure analysis revealed multiple beta-pleated sheets in hydrophilic clusters, providing a possible explanation for the immunogenic properties of the polypeptide.

摘要

人巨细胞病毒颗粒在其基质中含有一种表观分子量为150,000的磷蛋白(pp150);在蛋白质印迹分析中,该蛋白与人抗血清反应特别强烈。通过用单特异性兔抗血清筛选噬菌体λgt11 cDNA表达文库,对pp150基因进行了定位。随后,将cDNA与包含人巨细胞病毒AD169株基因组的黏粒和质粒克隆进行杂交,将该基因定位到HindIII片段J和N上。该基因转录成一个6.2千碱基的晚期RNA。测定了该区域的核苷酸序列,并通过引物延伸和核酸酶保护实验确定了一个丰富转录本的转录起始位点和两个聚腺苷酸化位点。通过计算机分析推导的pp150阅读框产生了一个长度为1048个氨基酸的多肽;蛋白质二级结构分析显示亲水区有多个β折叠片层,这为该多肽的免疫原性特性提供了一种可能的解释。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db1d/254110/156f84d8b525/jvirol00096-0069-a.jpg

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