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P1、P7和F微型质粒的配分函数。

The partition functions of P1, P7, and F miniplasmids.

作者信息

Austin S, Abeles A

出版信息

Basic Life Sci. 1985;30:215-26. doi: 10.1007/978-1-4613-2447-8_18.

Abstract

The partition regions of P1, P7, and F miniplasmids are discrete DNA sequences of about 3 kb in length that will promote accurate partition of hybrid plasmids independent of the source of replication functions or the position or orientation of the elements. Each of the par regions seems to be very similarly organized, with open reading frames for essential proteins and a terminal site which appears to be analogous to the centromere of eukaryotic cells. When cloned, these terminal sites exert incompatibility against their respective parent plasmids presumably because they can compete with the parent plasmids as substrates for partition. We have determined the complete DNA sequence of the P1 par region. In addition to the open reading frame for the essential parA protein (42-44 kd), the region contains a second open reading frame which could encode a 38-kd protein. The 2 large open reading frames appear to form an operon that is negatively regulated from a site adjacent to the promoter and responds to the par gene products in trans. Both this site and the downstream "centromere" site, incB, contain blocks of extremely AT-rich sequences, which are postulated to be binding sites for par proteins. The incB and upstream AT-rich regions both contain 20-bp imperfect inverted repeats. Further downstream from the minimal incB sequence (172 bp) lies an additional region which is essential for partition. The further analysis of the P1 par region should be greatly facilitated by the finding that it can function in cis to stabilize pBR322 vectors under conditions where the copy number of pBR322 is reduced.

摘要

P1、P7和F微型质粒的分配区域是长度约为3 kb的离散DNA序列,它们能够促进杂交质粒的准确分配,而与复制功能的来源或元件的位置及方向无关。每个par区域的组织方式似乎非常相似,都有必需蛋白质的开放阅读框和一个末端位点,该末端位点似乎类似于真核细胞的着丝粒。当被克隆时,这些末端位点会与其各自的亲本质粒产生不相容性,大概是因为它们可以作为分配的底物与亲本质粒竞争。我们已经确定了P1 par区域的完整DNA序列。除了必需的parA蛋白(42 - 44 kd)的开放阅读框外,该区域还包含另一个开放阅读框,它可能编码一种38 kd的蛋白质。这两个大的开放阅读框似乎形成了一个操纵子,该操纵子受到启动子附近一个位点的负调控,并对反式的par基因产物做出反应。这个位点和下游的“着丝粒”位点incB都包含富含AT的序列块,据推测这些序列块是par蛋白的结合位点。incB和上游富含AT的区域都包含20 bp的不完全反向重复序列。在最小的incB序列(172 bp)的更下游有一个对分配至关重要的额外区域。发现P1 par区域在顺式作用下能够在pBR322拷贝数减少的条件下稳定pBR322载体,这将极大地促进对P1 par区域的进一步分析。

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