哈维氏弧菌荧光素酶基因的克隆：合成寡核苷酸探针的应用。

Cohn D H, Ogden R C, Abelson J N, Baldwin T O, Nealson K H, Simon M I, Mileham A J

Proc Natl Acad Sci U S A. 1983 Jan;80(1):120-3. doi: 10.1073/pnas.80.1.120.

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.

使用混合序列合成寡核苷酸探针从哈维弧菌中分离出一个克隆，该克隆包含编码细菌荧光素酶α亚基的基因以及编码β亚基的部分基因。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。对两个结构基因之前区域的序列比较揭示了相当高的同源性，并确定了可能参与基因表达的位点。从该生物体的总基因组DNA克隆库中鉴定出一个克隆，这表明只要有该基因产物的一些蛋白质序列，混合探针就可以成功用于从任何细菌中分离出感兴趣的基因。