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从革兰氏阴性细菌黄粘球菌中分离出的多拷贝单链DNA。

Multicopy single-stranded DNA isolated from a gram-negative bacterium, Myxococcus xanthus.

作者信息

Yee T, Furuichi T, Inouye S, Inouye M

出版信息

Cell. 1984 Aug;38(1):203-9. doi: 10.1016/0092-8674(84)90541-5.

Abstract

A gram-negative bacterium, Myxococcus xanthus, was found to contain 500 to 700 copies per chromosome of a short single-stranded linear DNA fragment. When this DNA (multicopy single-stranded DNA; msDNA) labeled at the 5' end with kinase was used as a probe against total chromosomal blots, it hybridized to unique high molecular weight bands, which were cloned and sequenced. Labeling of msDNA was also possible using the Klenow fragment of DNA polymerase I as well as terminal deoxynucleotidyl transferase, permitting direct sequencing. The 5' end of msDNA was found to be primed by a short RNA segment. The DNA portion of msDNA consisted of 163 bases. Exact correspondence was seen between the msDNA sequence and the sequence of a chromosomal clone. An elaborate secondary structure is postulated for the msDNA sequence. A similar satellite DNA was also found in another myxobacterium, Stigmatella aurantiaca.

摘要

人们发现,革兰氏阴性细菌黄色黏球菌(Myxococcus xanthus)每条染色体含有500至700个短单链线性DNA片段拷贝。当用激酶在5'端标记的这种DNA(多拷贝单链DNA;msDNA)作为探针与总染色体印迹杂交时,它与独特的高分子量条带杂交,这些条带被克隆并测序。使用DNA聚合酶I的Klenow片段以及末端脱氧核苷酸转移酶也可以对msDNA进行标记,从而实现直接测序。发现msDNA的5'端由一个短RNA片段引发。msDNA的DNA部分由163个碱基组成。msDNA序列与染色体克隆序列之间存在精确对应关系。推测msDNA序列具有复杂的二级结构。在另一种黏细菌橙色标桩菌(Stigmatella aurantiaca)中也发现了类似的卫星DNA。

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