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细菌修饰甲基化酶基因在酵母中的表达。

Expression of a bacterial modification methylase gene in yeast.

作者信息

Fehér Z, Kiss A, Venetianer P

出版信息

Nature. 1983;302(5905):266-8. doi: 10.1038/302266a0.

Abstract

Methylation of specific cytosines in the DNA is generally believed to play some role in the regulation of gene expression in eukaryotes. However, some eukaryotes, such as Drosophila and yeast (S. Hattman, personal communication) seem not to contain 5-methylcytosine in their DNA. It would be interesting to test, how gene expression in such organisms would respond to the methylation of specific cytosines in the genome. As a first step towards this goal, we have introduced the gene encoding the Bacillus sphaericus R modification methylase, which methylates the internal cytosine within the recognition sequence 5'-GGCC, into yeast cells. Southern-type hybridization to DNAs isolated from the transformed yeast clones revealed that the yeast plasmid carrying the prokaryotic methylase gene, as well as the two chromosomal genes tested (his3 and leu2) were methylated, whereas the bulk of the yeast DNA remained largely unmethylated. This indicates that the Bacillus sphaericus modification methylase was expressed in yeast but it modified only certain parts of the yeast DNA.

摘要

一般认为,DNA中特定胞嘧啶的甲基化在真核生物基因表达调控中发挥一定作用。然而,一些真核生物,如果蝇和酵母(S. Hattman,个人交流),其DNA中似乎不含5-甲基胞嘧啶。测试此类生物中的基因表达如何响应基因组中特定胞嘧啶的甲基化将是一件有趣的事情。作为朝着这个目标迈出的第一步,我们已将编码球形芽孢杆菌R修饰甲基化酶的基因导入酵母细胞,该甲基化酶可使识别序列5'-GGCC内的内部胞嘧啶甲基化。对从转化的酵母克隆中分离出的DNA进行Southern型杂交分析表明,携带原核甲基化酶基因的酵母质粒以及测试的两个染色体基因(his3和leu2)发生了甲基化,而大部分酵母DNA在很大程度上仍未甲基化。这表明球形芽孢杆菌修饰甲基化酶在酵母中表达,但它仅修饰酵母DNA的某些部分。

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