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肺炎球菌的转化:隐蔽期复合体中脱氧核糖核酸的核酸酶抗性

Transformation in pneumococcus: nuclease resistance of deoxyribonucleic acid in the eclipse complex.

作者信息

Morrison D A, Mannarelli B

出版信息

J Bacteriol. 1979 Nov;140(2):655-65. doi: 10.1128/jb.140.2.655-665.1979.

Abstract

Donor deoxyribonucleic acid strands in the eclipse phase of genetic transformation of pnuemococcus (Streptococcus pneumoniae) are purified as a complex with a cf the deoxyribonucleic acid strand in this complex to digestion by nucleases was shown to be 50- to 1,000-fold less than that of uncomplexed single strands of deoxyribonucleic acid. Deoxyribonuclease I, micrococcal nuclease, Neurospora endonuclease, nuclease P1, and the major endogenous nuclease of cell-free extracts were studied. Sensitivity to nuclease attack was not uniform along the deoxyribonucleic acid strand; sequences of strongly protected bases were separated by more sensitive regions. The minimum size of protected fragments was about 70 bases. A complex of protein with the protected deoxyribonucleic acid segments was obtained after partial digestion. The sizes of these complexes, of the protected deoxyribonucleic acid segments, and of the protein subunit released by complete nuclease digestion, are all approximately identical, as determined by gel exclusion chromatography. Deoxyribonucleic acid strands of eclipse complex were also shown to be particularly well protected from attack by the major pneumococcal endonuclease in cell extracts.

摘要

在肺炎球菌(肺炎链球菌)遗传转化的隐蔽期,供体脱氧核糖核酸链作为一种复合物被纯化,该复合物中的脱氧核糖核酸链对核酸酶消化的敏感性比未复合的单链脱氧核糖核酸低50至1000倍。研究了脱氧核糖核酸酶I、微球菌核酸酶、脉孢菌核酸酶、核酸酶P1以及无细胞提取物中的主要内源性核酸酶。沿着脱氧核糖核酸链,对核酸酶攻击的敏感性并不均匀;受到强烈保护的碱基序列被更敏感的区域隔开。受保护片段的最小尺寸约为70个碱基。部分消化后获得了蛋白质与受保护的脱氧核糖核酸片段的复合物。通过凝胶排阻色谱法测定,这些复合物、受保护的脱氧核糖核酸片段以及核酸酶完全消化后释放的蛋白质亚基的大小都大致相同。还表明,隐蔽期复合物的脱氧核糖核酸链特别能抵御细胞提取物中主要肺炎球菌内切核酸酶的攻击。

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