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形态不同的噬菌体LP 52和θ的基因组之间的序列同源性与重组

Sequence homology and recombination between the genomes of morphologically dissimilar bacteriophages LP 52 and theta.

作者信息

Forstová J, Grünnerová H, Hostomský Z, Doskocil J

出版信息

Mol Gen Genet. 1982;187(1):138-47. doi: 10.1007/BF00384397.

Abstract

A restriction fragment map of Bacillus licheniformis temperate phage LP 52 DNA (molecular weight 38.5 X 10(6)) was established, using restriction endonucleases BamHI (8 target sites), BglI (10 sites,) BglII (13 sites) and EcoRI (22 sites). The map is linear, with well-defined ends, without any signs of circular permutation. The DNA of a related phage, LP 51, produced identical restriction fragments. At least 62% DNA of LP 52 has been found homologous to the DNA of the recently discovered, morphologically quite dissimilar, phage I, as demonstrated by hybridization of electrophoretically separated restriction fragments of DNA. Under the same conditions, the DNAs of LP 52 and of the morphologically similar Bacillus subtilis phage phi 105 did not cross-hybridize. The homologous regions in the genomes of phages LP 52 and I have been shown to be colinear. Comparison of the cleavage maps of phages LP 52 and I has shown that, within the regions of homology, not a single restriction fragment and few restriction sites have been conserved during divergent evolution. Three major regions of heterology were defined; the longest one, covering the right-hand end of the map (73 +/- 2.75% up to 100% LP 52 genome length) appeared to contain genes coding for structural proteins of the virions; a shorter region at the left-hand end of the map (coordinates zero to 10.3 +/- 3.3% LP 52 genome length) and a very short central region (coordinates 41.8-43.9%) could be identified, the latter apparently containing a regulatory locus responsible for the heteroimmune behavior of the two phages. Recombinants between phages LP 52 and I were isolated. Mapping of recombinant genomes has indicated mutual substitution of allelic pieces of LP 52 and I DNAs upon strict conservation of overall genome length.

摘要

利用限制性内切酶BamHI(8个靶位点)、BglI(10个位点)、BglII(13个位点)和EcoRI(22个位点)构建了地衣芽孢杆菌温和噬菌体LP 52 DNA(分子量38.5×10⁶)的限制性片段图谱。该图谱是线性的,末端明确,没有任何环状排列的迹象。相关噬菌体LP 51的DNA产生了相同的限制性片段。通过对DNA电泳分离的限制性片段进行杂交证明,LP 52至少62%的DNA与最近发现的、形态上差异很大的噬菌体I的DNA同源。在相同条件下,LP 52和形态相似的枯草芽孢杆菌噬菌体phi 105的DNA没有交叉杂交。噬菌体LP 52和I基因组中的同源区域已被证明是共线的。对噬菌体LP 52和I的切割图谱比较表明,在同源区域内,在分歧进化过程中没有一个限制性片段和很少的限制性位点被保留下来。确定了三个主要的异源区域;最长的一个覆盖图谱的右端(占LP 52基因组长度的73±2.75%至100%),似乎包含编码病毒粒子结构蛋白的基因;在图谱左端的一个较短区域(坐标为零至占LP 52基因组长度的10.3±3.3%)和一个非常短的中央区域(坐标为41.8 - 43.9%)可以被识别,后者显然包含一个负责这两种噬菌体异免疫行为的调控位点。分离到了噬菌体LP 52和I之间的重组体。重组基因组的图谱表明,在严格保持总基因组长度的情况下,LP 52和I的DNA等位片段相互替换。

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