THE ISOLATION AND CHARACTERIZATION OF THE ESCHERICHIA-COLI DNA ADENINE METHYLASE (DAM) GENE
THE ISOLATION AND CHARACTERIZATION OF THE ESCHERICHIA-COLI DNA ADENINE METHYLASE (DAM) GENE
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DOI：
10.1093/nar/11.3.837
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发表时间：
1983-01-01
影响因子：
14.9
通讯作者：
GINGERAS, TR
中科院分区：
文献类型：
--
作者：
BROOKS, JE;BLUMENTHAL, RM;GINGERAS, TR
已知Ecolidam（DNA腺嘌呤甲基化酶）烯炔使序列GATC甲基化。用序列特异性DNA甲基化酶基因克隆的一般方法，将dam基因克隆到1.14kb的片段上，插入质粒载体pBK 322中。随后的限制性作图和亚克隆实验建立了一组基因的近似边界。dju基因的核苷酸序列被确定，对该序列的分析揭示了一个独特的开放阅读框架，其长度与编码dam的六个蛋白质所必需的长度相对应。从该序列得到的氨基酸组成与纯化的蛋白质的氨基酸组成密切对应。用酶切和DNA：DNA杂交方法研究了dasi基因在多种原核生物中的可能存在。
The Ecolidam (DNA adenine methylase) enxyne is known to nethylate the sequence GATC. A general nethod for cloning sequence-speoific DNA methylase genet was used to isolate thedamgene on a 1.14 kb fragment, inserted in the plasmid vector pBK322. Subsequent restriction mapping and subcloning experiments established a set of approximate boundaries of the gene. The nucleotide sequence of the dju gene was determined, and analysis of that sequence revealed a unique open reading frame which corresponded in length to that necessary to oode for a protein the sixe ofdam. Amino acid conposition derived froa this sequence corresponds closely to the amino acid composition of the purifieddamprotein. Enxynatic and DNA:DNA hybridixation methods were used to investigate the possible presence of dasi genes in a variety of prokaryotlc organisms.