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噬菌体T5 DNA切口和末端的序列分析

Sequence analysis of the nicks and termini of bacteriophage T5 DNA.

作者信息

Nichols B P, Donelson J E

出版信息

J Virol. 1977 May;22(2):520-6. doi: 10.1128/JVI.22.2.520-526.1977.

Abstract

Bacteriophage T5 DNA, when isolated from mature phage particles, contains several nicks in one of the two strands. The 5'-terminal nucleotides at the nicks were labeled with polynucleotide kinase and [gamma-32P]ATP, and the 3'-terminal nucleotides were labeled with Escherichia coli DNA polymerase I and [alpha-32P]dGTP. The sequences around the nicks were analyzed by partial nuclease digestion followed by homochromatography fractionation of the resulting oligonucleotides. The nicks had at least the sequence -PuOH pGpCpGpC- in common. In addition, the two 5' external termini had the first seven nucleotides in common.

摘要

从成熟噬菌体颗粒中分离出的噬菌体T5 DNA,在两条链中的一条链上含有几个切口。用多核苷酸激酶和[γ-32P]ATP标记切口处的5'-末端核苷酸,用大肠杆菌DNA聚合酶I和[α-32P]dGTP标记3'-末端核苷酸。通过部分核酸酶消化,然后对所得寡核苷酸进行同系层析分级分离,分析切口周围的序列。这些切口至少有-PuOH pGpCpGpC-的序列。此外,两个5'外部末端有相同的前七个核苷酸。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf5e/515742/b0292d549a5e/jvirol00209-0292-a.jpg

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