Promoter cloning in the radioresistant bacterium Deinococcus radiodurans
Promoter cloning in the radioresistant bacterium Deinococcus radiodurans
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DOI：
10.1128/jb.183.10.3169-3175.2001
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发表时间：
2001-05-01
影响因子：
3.2
通讯作者：
Lidstrom, ME
中科院分区：
文献类型：
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作者：
Meima, R;Rothfuss, HM;Lidstrom, ME
耐辐射异常球菌是一种高度耐辐射的细菌，属于细菌结构域的一个主要分支。由于对这种细菌中的基因表达知之甚少，因此对启动子进行了初步研究。为了分离启动子和研究启动子的功能，我们构建了一系列整合型载体。开发了抗辐射剂。这些载体是基于大肠杆菌复制子，不能自主复制的D，radiodurans和携带同源序列的D，radiodurans染色体中的置换重组，所得到的整合载体被用来研究报告基因的表达融合到一些推定的启动子，从D，radiodurans R1基因组扩增。通过北方杂交和引物延伸实验对这些和其它推定的启动子进行进一步分析。与以前的报道相反，这些启动子的-10和-35区域类似于大肠杆菌的sigma（70）共有序列，杆菌
Deinococcus radiodurans is a highly radiation-resistant bacterium that is classed in a major subbranch of the bacterial domain. Since very little is known about gene expression in this bacterium, an initial study of promoters was undertaken. In order to isolate promoters and study promoter function, a series of integrative vectors for stable chromosomal insertion in D. radiodurans were developed. These vectors are based on Escherichia coli replicons that are unable to replicate autonomously in D, radiodurans and carry homologous sequences for replacement recombination in the D, radiodurans chromosome, The resulting integration vectors were used to study expression of reporter genes fused to a number of putative promoters that were amplified from the D, radiodurans R1 genome. Further analysis of these and other putative promoters was performed by Northern hybridization and primer extension experiments. In contrast to previous reports, the -10 and -35 regions of these promoters resembled the sigma (70) consensus sequence of E., coli.