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编码病毒DNA依赖性RNA聚合酶30千道尔顿亚基的痘苗病毒基因。

Vaccinia virus gene encoding a 30-kilodalton subunit of the viral DNA-dependent RNA polymerase.

作者信息

Broyles S S, Pennington M J

机构信息

Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907-6799.

出版信息

J Virol. 1990 Nov;64(11):5376-82. doi: 10.1128/JVI.64.11.5376-5382.1990.

Abstract

Antibody was raised against purified vaccinia virus RNA polymerase and used to screen a recombinant vaccinia virus-lambda gt11 library. The DNA from several immunopositive clones was shown by Southern hybridization to originate from the vaccinia virus HindIII E fragment. The nucleotide sequence of the RNA polymerase subunit gene predicts a polypeptide 287 amino acids in length and 30,000 daltons in mass. An early transcript with a 5' terminus just upstream of the putative initiation codon was identified by S1 nuclease protection and primer extension analyses, demonstrating that this RNA polymerase subunit is expressed as an early viral gene product. The RNA polymerase subunit was synthesized by a bacterial expression vector to demonstrate that it corresponds to the previously described 37,000-dalton RNA polymerase subunit.

摘要

制备了针对纯化痘苗病毒RNA聚合酶的抗体,并用于筛选重组痘苗病毒-λgt11文库。通过Southern杂交显示,几个免疫阳性克隆的DNA源自痘苗病毒HindIII E片段。RNA聚合酶亚基基因的核苷酸序列预测其编码的多肽长度为287个氨基酸,质量为30,000道尔顿。通过S1核酸酶保护和引物延伸分析鉴定出一个5'末端恰好在推定起始密码子上游的早期转录本,表明该RNA聚合酶亚基作为早期病毒基因产物表达。该RNA聚合酶亚基由细菌表达载体合成,以证明它对应于先前描述的37,000道尔顿的RNA聚合酶亚基。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d74c/248587/9bd5864cdfd3/jvirol00066-0155-a.jpg

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