粟酒裂殖酵母RNA聚合酶II亚基2编码基因rpb2的克隆与序列测定。
Cloning and sequence determination of the Schizosaccharomyces pombe rpb2 gene encoding the subunit 2 of RNA polymerase II.
作者信息
Kawagishi M, Yamagishi M, Ishihama A
机构信息
Department of Molecular Genetics, National Institute of Genetics, Shizuoka, Japan.
出版信息
Nucleic Acids Res. 1993 Feb 11;21(3):469-73. doi: 10.1093/nar/21.3.469.
The gene, rpb2, encoding the second largest subunit, subunit 2, of RNA polymerase II has been cloned from Schizosaccharomyces pombe using the corresponding gene, RPB2, of Saccharomyces cerevisiae as a probe for cross-hybridization. We have determined the complete nucleotide sequence of rpb2, and parts of the PCR-amplified rpb2 cDNA. The predicted coding sequence of a polypeptide of 1210 amino acid residues with a calculated molecular weight of 138 kilodaltons was interrupted by a short intron. The overall amino acid sequence homology of the S. pombe subunit 2 is 68, 62 and 62% with the corresponding protein from S. cerevisiae, D. melanogaster and H. sapiens, respectively. Southern analysis of the genomic DNA digested with various restriction enzymes showed that rpb2 was present as a single copy in the S. pombe genome. Northern analysis showed that the transcript of rpb2 was about 4 kb in length.
利用酿酒酵母的相应基因RPB2作为交叉杂交探针，从粟酒裂殖酵母中克隆出编码RNA聚合酶II第二大亚基（亚基2）的基因rpb2。我们已经确定了rpb2的完整核苷酸序列以及PCR扩增的rpb2 cDNA的部分序列。预测的编码1210个氨基酸残基的多肽的编码序列被一个短内含子打断，其计算分子量为138千道尔顿。粟酒裂殖酵母亚基2与酿酒酵母、黑腹果蝇和智人的相应蛋白质的总体氨基酸序列同源性分别为68%、62%和62%。用各种限制酶消化基因组DNA的Southern分析表明，rpb2在粟酒裂殖酵母基因组中以单拷贝形式存在。Northern分析表明，rpb2的转录本长度约为4kb。
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