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噬菌体phi 29末端蛋白DNA连接位点的定点诱变：Ser232→Thr突变体的分离与鉴定

Site-directed mutagenesis in the DNA linking site of bacteriophage phi 29 terminal protein: isolation and characterization of a Ser232----Thr mutant.

作者信息

Garmendia C, Salas M, Hermoso J M

机构信息

Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma, Madrid, Spain.

出版信息

Nucleic Acids Res. 1988 Jul 11;16(13):5727-40. doi: 10.1093/nar/16.13.5727.

DOI:10.1093/nar/16.13.5727
PMID:3135531
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC336825/
Abstract

By site-directed mutagenesis we have changed the serine residue 232 of the phi 29 terminal protein, involved in the covalent linkage to dAMP for the initiation of replication, into a threonine residue. The mutant terminal protein has been purified to homogeneity and shown to be inactive in the formation of the initiation complex; nevertheless, the mutant protein retains its ability to interact with the phi 29 DNA polymerase and with the DNA. The results obtained indicate a high specificity in the linking site of the terminal protein.

摘要

通过定点诱变，我们已将参与与dAMP共价连接以启动复制的φ29末端蛋白的丝氨酸残基232替换为苏氨酸残基。突变的末端蛋白已被纯化至同质，并显示在起始复合物形成中无活性；然而，突变蛋白保留了其与φ29 DNA聚合酶和DNA相互作用的能力。所得结果表明末端蛋白的连接位点具有高度特异性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/7f9da961a299/nar00156-0036-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/2db941116556/nar00156-0033-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/a6098508960f/nar00156-0034-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/a1d4023d9eae/nar00156-0035-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/7f9da961a299/nar00156-0036-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/2db941116556/nar00156-0033-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/a6098508960f/nar00156-0034-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/a1d4023d9eae/nar00156-0035-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c444/336825/7f9da961a299/nar00156-0036-a.jpg

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Site-directed mutagenesis in the DNA linking site of bacteriophage phi 29 terminal protein: isolation and characterization of a Ser232----Thr mutant.噬菌体phi 29末端蛋白DNA连接位点的定点诱变：Ser232→Thr突变体的分离与鉴定
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本文引用的文献

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Initiation of phage phi 29 DNA replication in vitro: formation of a covalent complex between the terminal protein, p3, and 5'-dAMP.噬菌体φ29 DNA体外复制的起始：末端蛋白p3与5'-dAMP之间共价复合物的形成。
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Nucleotide sequence of the major early region of bacteriophage phi 29.噬菌体φ29主要早期区域的核苷酸序列。
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Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination.
揭示枯草芽孢杆菌中病毒组蛋白样蛋白的体内组织，由其识别病毒基因组的能力介导。
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Reverse transcription in hepatitis B viruses is primed by a tyrosine residue of the polymerase.乙型肝炎病毒中的逆转录由聚合酶的一个酪氨酸残基引发。
J Virol. 1994 Jan;68(1):6-13. doi: 10.1128/JVI.68.1.6-13.1994.
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Functional domains in the bacteriophage phi 29 terminal protein for interaction with the phi 29 DNA polymerase and with DNA.噬菌体φ29末端蛋白中与φ29 DNA聚合酶及DNA相互作用的功能结构域。
Nucleic Acids Res. 1989 Dec 25;17(24):10353-66. doi: 10.1093/nar/17.24.10353.
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Mutations of the precursor to the terminal protein of adenovirus serotypes 2 and 5.腺病毒血清型2和5末端蛋白前体的突变
J Virol. 1989 Dec;63(12):5244-50. doi: 10.1128/JVI.63.12.5244-5250.1989.
缓冲液梯度凝胶和35S标记辅助快速DNA序列测定。
Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963-5. doi: 10.1073/pnas.80.13.3963.
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Replication of bacteriophage phi 29 DNA in vitro: the roles of terminal protein and DNA polymerase.噬菌体φ29 DNA的体外复制：末端蛋白和DNA聚合酶的作用。
Proc Natl Acad Sci U S A. 1984 Sep;81(17):5374-8. doi: 10.1073/pnas.81.17.5374.
5
Characterization and purification of a phage phi 29-encoded DNA polymerase required for the initiation of replication.用于复制起始的噬菌体φ29编码的DNA聚合酶的表征与纯化。
Proc Natl Acad Sci U S A. 1984 Sep;81(17):5325-9. doi: 10.1073/pnas.81.17.5325.
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Purification in a functional form of the terminal protein of Bacillus subtilis phage phi 29.枯草芽孢杆菌噬菌体phi 29末端蛋白的功能形式纯化
Proc Natl Acad Sci U S A. 1984 Mar;81(6):1639-43. doi: 10.1073/pnas.81.6.1639.
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High level synthesis in Escherichia coli of the Bacillus subtilis phage phi 29 proteins p3 and p4 under the control of phage lambda PL promoter.在噬菌体λ PL 启动子的控制下，枯草芽孢杆菌噬菌体 phi 29 蛋白 p3 和 p4 在大肠杆菌中的高水平合成。
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Nucleic Acids Res. 1981 Jan 24;9(2):309-21. doi: 10.1093/nar/9.2.309.