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HincII限制修饰系统的克隆、核苷酸序列及表达

Cloning, nucleotide sequence, and expression of the HincII restriction-modification system.

作者信息

Ito H, Sadaoka A, Kotani H, Hiraoka N, Nakamura T

机构信息

Bioproducts Development Center, Takara Shuzo Co., Ltd., Shiga, Japan.

出版信息

Nucleic Acids Res. 1990 Jul 11;18(13):3903-11. doi: 10.1093/nar/18.13.3903.

DOI:10.1093/nar/18.13.3903
PMID:2374714
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC331092/
Abstract

Two genes, coding for the HincII from Haemophilus influenzae Rc restriction-modification system, were cloned and expressed in Escherichia coli RR1. Their DNA sequences were determined. The HincII methylase (M.HincII) gene was 1,506 base pairs (bp) long, corresponding to a protein of 502 amino acid residues (Mr = 55,330). The HincII endonuclease (R.HincII) gene was 774 bp long, corresponding to a protein of 258 amino acid residues (Mr = 28,490). The amino acid residues predicted from the R.HincII and the N-terminal amino acid sequence of the enzyme found by analysis were identical. These methylase and endonuclease genes overlapped by 1 bp on the H. influenzae Rc chromosomal DNA. The clone, named E. coli RR1-Hinc, overproduced R.HincII. The R.HincII activity of this clone was 1,000-fold that from H. influenzae Rc. The amino acid sequence of M.HincII was compared with the sequences of four other adenine-specific type II methylases. Important homology was found between tne M.HincII and these other methylases.

摘要

编码流感嗜血杆菌Rc限制修饰系统中HincII的两个基因被克隆并在大肠杆菌RR1中表达。测定了它们的DNA序列。HincII甲基化酶（M.HincII）基因长1506个碱基对（bp），对应于一个由502个氨基酸残基组成的蛋白质（Mr = 55330）。HincII核酸内切酶（R.HincII）基因长774 bp，对应于一个由258个氨基酸残基组成的蛋白质（Mr = 28490）。通过分析预测的R.HincII氨基酸残基与所发现的该酶的N端氨基酸序列相同。这些甲基化酶和核酸内切酶基因在流感嗜血杆菌Rc染色体DNA上重叠1 bp。该克隆名为大肠杆菌RR1-Hinc，过量产生R.HincII。该克隆的R.HincII活性是流感嗜血杆菌Rc的1000倍。将M.HincII的氨基酸序列与其他四种腺嘌呤特异性II型甲基化酶的序列进行了比较。在M.HincII与这些其他甲基化酶之间发现了重要的同源性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/120e/331092/04ba0c38228b/nar00197-0216-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/120e/331092/9a5e4f07e320/nar00197-0213-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/120e/331092/04ba0c38228b/nar00197-0216-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/120e/331092/9a5e4f07e320/nar00197-0213-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/120e/331092/04ba0c38228b/nar00197-0216-a.jpg

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