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奇异变形杆菌裂解物快速沉降部分中F'质粒和R质粒脱氧核糖核酸的差异关联

Differential association of F' plasmid and R plasmid deoxyribonucleic acid with a rapidly sedimenting fraction of a Proteus mirabilis lysate.

作者信息

Taichman L, Rownd R H

出版信息

J Bacteriol. 1977 Jun;130(3):1262-73. doi: 10.1128/jb.130.3.1262-1273.1977.

Abstract

We have examined the association of an F' plasmid and an R plasmid in Proteus mirabilis with a rapidly sedimenting material that is generated by sodium dodecyl sulfate lysis and low speed centrifugation. Virtually all of the chromosomal deoxyribonucleic acid (DNA) and the F' plasmid DNA are associated with the rapidly sedimenting material after gentle lysis and centrifugation. A portion of R plasmid NR1 DNA (usually 5 to 25%) is not bound to the rapidly sedimenting material and is recovered in the supernatant fraction. This difference in binding is not related to the size of the plasmid DNA, since F' plasmids and R plasmids of different molecular weights showed the same behavior. R plasmid DNA labeled by a brief pulse of [(3)H]thymine is recovered in the supernatant fraction to a lower extent than the total R plasmid DNA. It would appear that R plasmid replication takes place in association with the rapidly sedimenting material. With prolongation of the [(3)H]thymine pulse, the [(3)H]thymine-labeled R plasmid DNA is recovered in the supernatant fraction with the same probability as the total R plasmid DNA. This finding indicates that a change in R plasmid attachment to the rapidly sedimenting material occurs some time after its replication. The differences observed in the replication of F' plasmids and R plasmids in P. mirabilis may be related to their different modes of association with the rapidly sedimenting material.

摘要

我们研究了奇异变形杆菌中F'质粒和R质粒与一种快速沉降物质的关联,该物质由十二烷基硫酸钠裂解和低速离心产生。经过温和裂解和离心后,几乎所有的染色体脱氧核糖核酸(DNA)和F'质粒DNA都与快速沉降物质相关联。一部分R质粒NR1 DNA(通常为5%至25%)不与快速沉降物质结合,而是在上清液部分中回收。这种结合差异与质粒DNA的大小无关,因为不同分子量的F'质粒和R质粒表现出相同的行为。经短暂脉冲[(3)H]胸腺嘧啶标记的R质粒DNA在上清液部分中的回收率低于总R质粒DNA。似乎R质粒复制与快速沉降物质相关联。随着[(3)H]胸腺嘧啶脉冲时间的延长,[(3)H]胸腺嘧啶标记的R质粒DNA在上清液部分中的回收率与总R质粒DNA相同。这一发现表明,R质粒与快速沉降物质的附着在其复制后的某个时间发生了变化。在奇异变形杆菌中观察到的F'质粒和R质粒复制的差异可能与其与快速沉降物质的不同关联方式有关。

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