A novel retron that produces RNA-less msDNA in Escherichia coli using reverse transcriptase.

A novel retron that produces RNA-less msDNA in Escherichia coli using reverse transcriptase.
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一种新型逆转录子,可使用逆转录酶在大肠杆菌中产生无 RNA msDNA。

DOI:
10.1006/plas.1997.1298
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发表时间:
1997
期刊:
影响因子:
2.6
通讯作者:
D. Lim
D. Lim
中科院分区:
生物学3区
文献类型:
--
作者:
T. Lima;D. Lim

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细菌逆转录因子,或称逆转录因子,利用逆转录酶(RT)产生与RNA共价连接的多拷贝单链DNA (msDNA)分子。在这些研究中,我们发现临床分离的大肠杆菌110的逆转录子产生一种具有5'磷酸残基的新型无rna msDNA。msDNA是一个74个核苷酸的单链DNA分子,具有稳定的茎环结构,没有错配的碱基对。只有编码msDNA (msd)、msdRNA (msr)和RT (ret)的基因才能产生msDNA分子。这些基因在逆转录子上的组织与其他产生支链msDNA-RNA的元件相似。保守的鸟嘌呤是msDNA-RNA复合物中的支链残基,是分支形成所必需的,也存在。定点突变表明,这种鸟嘌呤是产生无rna msDNA所必需的。我们假设菌株110中无rna的msDNA是由支链msDNA- rna化合物的核裂解产生的。
Bacterial retroelements, or retrons, use reverse transcriptase (RT) to produce a multicopy single-stranded DNA (msDNA) molecule that is covalently linked to RNA. In these studies we show that a retron from Escherichia coli 110, a clinical isolate, produces a novel RNA-less msDNA with a 5' phosphate residue. The msDNA is a 74-nucleotide single-stranded DNA molecule with a stable stem-loop structure without a mismatched base pair. Only the genes encoding msDNA (msd), msdRNA (msr), and RT (ret) are required to produce the msDNA molecule. The organization of these genes on the retron was similar to that of other elements producing branched msDNA-RNA. The conserved guanine, which is the branched residue in msDNA-RNA complexes and is essential for branch formation, is also present. Site-directed mutagenesis showed that this guanine is essential for the production of RNA-less msDNA. We postulate that the RNA-less msDNA in strain 110 is produced by nucleolytic cleavage of the branched msDNA-RNA compound.
DOI: --
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