Dolganov N, Grossman A R
Department of Plant Biology, Carnegie Institution of Washington, Stanford, California 94305.
J Bacteriol. 1993 Dec;175(23):7644-51. doi: 10.1128/jb.175.23.7644-7651.1993.
We have developed a simple procedure for generating mutants of the cyanobacterium Synechococcus sp. strain PCC 7942 in which the site of the lesion can be readily identified. This procedure involves transforming Synechococcus sp. strain PCC 7942 with a library of its own DNA that was fully digested with Sau3A and ligated into the plasmid vector pUC8. The homologous integration of the recombinant plasmid into the genome will often result in the disruption of a gene and the loss of gene function. We have used this method to generate many mutants of Synechococcus sp. strain PCC 7942 which grow as multicellular filaments rather than as unicells. Since the gene harboring the lesion was tagged with pUC8, it was easily isolated. In this paper, we discuss the usefulness of this procedure for the generation of mutants, and we characterize one mutant in which the lesion may be in an operon involved in the assembly of lipopolysaccharides.
我们已经开发出一种简单的方法来生成蓝藻聚球藻属（Synechococcus sp.）菌株PCC 7942的突变体，其中损伤位点能够很容易地被识别出来。该方法包括用经Sau3A完全消化并连接到质粒载体pUC8中的聚球藻属菌株PCC 7942自身的DNA文库来转化聚球藻属菌株PCC 7942。重组质粒与基因组的同源整合常常会导致基因的破坏和基因功能的丧失。我们已经使用这种方法生成了许多聚球藻属菌株PCC 7942的突变体，这些突变体以多细胞丝状体而非单细胞的形式生长。由于携带损伤的基因用pUC8进行了标记，所以很容易将其分离出来。在本文中，我们讨论了该方法在生成突变体方面的实用性，并对一个突变体进行了表征，该突变体中的损伤可能存在于一个参与脂多糖组装的操纵子中。
