Gay P, Le Coq D, Steinmetz M, Ferrari E, Hoch J A
J Bacteriol. 1983 Mar;153(3):1424-31. doi: 10.1128/jb.153.3.1424-1431.1983.
A clone bearing the structural gene sacB, coding for the exoenzyme levansucrase, was isolated from a library of Bacillus subtilis DNA that was cloned in phage lambda charon 4A on the basis of the transforming activity of the chimeric DNA. This lambda clone also was found to contain the sacR and smo loci. Subcloning the sacB-sacR region in plasmid pBR325 resulted in a clone which directed levansucrase synthesis in Escherichia coli. The nucleotide sequence coding for the secreted protein was localized on the physical map of the cloned DNA.
从枯草芽孢杆菌DNA文库中分离出一个携带编码外切酶果聚糖蔗糖酶的结构基因sacB的克隆。该文库是基于嵌合DNA的转化活性克隆到噬菌体λ载体Charon 4A中的。还发现这个λ克隆含有sacR和smo基因座。将sacB - sacR区域亚克隆到质粒pBR325中,得到一个能在大肠杆菌中指导果聚糖蔗糖酶合成的克隆。编码分泌蛋白的核苷酸序列定位在克隆DNA的物理图谱上。