红平红球菌噬菌体NJL基因组的特征分析

Characterization of the genome of the Rhodococcus rhodochrous bacteriophage NJL.

作者信息

Sunairi M, Watanabe T, Oda H, Murooka H, Nakajima M

机构信息

Department of Applied Biological Science, College of Agriculture and Veterinary Science, Nihon University, Kanagawa, Japan.

出版信息

Appl Environ Microbiol. 1993 Jan;59(1):97-100. doi: 10.1128/aem.59.1.97-100.1993.

DOI:10.1128/aem.59.1.97-100.1993

PMID:8439171

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC202061/

Abstract

Temperate bacteriophage NJL of Rhodococcus rhodochrous has a 49-kb linear double-stranded DNA with cohesive ends (cos). NJL DNA has unique target sites for HindIII and SspI, two target sites each for NheI and ScaI, and no cleavage site for AxyI, DraI, EcoRI, SacI, and SphI. The single-stranded regions of cos ends were ligated to each other with T4 DNA ligase, removed with mung bean nuclease, or blunted with the Klenow large fragment of DNA polymerase I; then the sequences of the cos ends were determined. Comparison of these sequences revealed that the single-stranded regions are complementary and 18 bases long and protrude at the 3' ends; they have the following sequences: 5'-TTGGCACCGTGGGAGGAG-3' and 3'-AACCGTGGCAC CCTCCTC-5'. A physical map of NJL was constructed by a cos mapping method based on information about the structure of the cohesive ends and multiple digestions with restriction endonucleases.

摘要

红平红球菌的温和噬菌体NJL具有一条49千碱基对的线性双链DNA，其末端带有粘性末端（cos）。NJL DNA对HindIII和SspI有独特的靶位点，对NheI和ScaI各有两个靶位点，对AxyI、DraI、EcoRI、SacI和SphI没有切割位点。cos末端的单链区域用T4 DNA连接酶相互连接，用绿豆核酸酶去除，或用DNA聚合酶I的Klenow大片段使其平端化；然后测定cos末端的序列。这些序列的比较显示，单链区域是互补的，长度为18个碱基，在3'端突出；它们具有以下序列：5'-TTGGCACCGTGGGAGGAG-3'和3'-AACCGTGGCAC CCTCCTC-5'。基于粘性末端结构信息和用限制性内切酶进行的多次消化，通过cos图谱法构建了NJL的物理图谱。