Cloning of the Vibrio harveyi luciferase genes: use of a synthetic oligonucleotide probe.
Cloning of the Vibrio harveyi luciferase genes: use of a synthetic oligonucleotide probe.
复制标题
哈维氏弧菌荧光素酶基因的克隆：使用合成寡核苷酸探针。
DOI：
--
复制
发表时间：
1983
影响因子：
11.1
通讯作者：
A. Mileham
中科院分区：
文献类型：
--
作者：
D. Cohn;R. Ogden;J. Abelson;T. Baldwin;K. Nealson;M. Simon;A. Mileham
使用混合序列合成寡核苷酸探针分离含有编码来自哈维氏弧菌的细菌荧光素酶α亚基的基因和编码β亚基的部分基因的克隆。 DNA序列分析使我们能够确定这些基因在细菌染色体上紧密相连并以相同方向转录。两个结构基因之前区域的序列比较揭示了相当大的同源性，并鉴定了可能参与基因表达的位点。从该生物体的总基因组 DNA 克隆库中鉴定克隆表明，只要可获得基因产物的某些蛋白质序列，混合探针就可以成功地从任何细菌中分离出感兴趣的基因。
A mixed-sequence synthetic oligonucleotide probe was used to isolate a clone containing the gene encoding the alpha subunit of bacterial luciferase from Vibrio harveyi and part of the gene coding for the beta subunit. DNA sequence analysis has allowed us to determine that the genes are closely linked on the bacterial chromosome and transcribed in the same direction. Comparison of the sequences in the regions preceding the two structural genes has revealed considerable homology and has identified sites that may be involved in the expression of the genes. Identification of a clone from a clone bank of total genomic DNA from this organism shows that mixed probes can be successfully used to isolate a gene of interest from any bacterium provided some protein sequence for the gene product is available.