Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts 02115; email:
Annu Rev Microbiol. 2013;67:1-19. doi: 10.1146/annurev-micro-092412-155732.
I recount the history of how I became interested in the use of gene fusions for studying biological problems. Initially, selections for mutations that would restore function to an inactivated lac operon unexpectedly yielded fusions in which lac was expressed from the controlling elements of upstream genes. Subsequently, by chance, I generated strains in which the lac operon was transposed from its normal position on the chromosome to a position close to the trp operon, thus facilitating sets of useful fusions of the two operons. The development of a more generalized technique for obtaining fusions by my student Malcolm Casadaban opened up a much broader set of biological problems that could be approached with fusions. Work on these problems included the study of protein translocation across membranes, the analysis of membrane protein topology, and the discovery of the pathway of electron transfer that leads to disulfide bond formation in proteins.
我讲述了自己如何对利用基因融合来研究生物学问题产生兴趣的历史。最初，为了筛选出能够恢复失活 lac 操纵子功能的突变体，我意外地得到了 lac 从上游基因调控元件表达的融合产物。随后，机缘巧合之下，我构建了 lac 操纵子从染色体正常位置易位到接近 trp 操纵子位置的菌株，从而促进了两个操纵子之间的一系列有用融合。我的学生马尔科姆·卡萨丹班（Malcolm Casadaban）开发了一种更通用的融合获取技术，为利用融合来解决更广泛的生物学问题开辟了道路。这些问题的研究包括跨膜蛋白转运、膜蛋白拓扑分析以及发现导致蛋白质中二硫键形成的电子转移途径。
