Ortlepp S A, Ollington J F, McConnell D J
Gene. 1983 Sep;23(3):267-76. doi: 10.1016/0378-1119(83)90017-3.
A resident-plasmid cloning system developed for Bacillus subtilis has been used to isolate recombinant plasmids carrying DNA from Bacillus licheniformis which confer alpha-amylase activity on alpha-amylase-negative mutants of B. subtilis. These plasmids contain a 3550-bp insert at the EcoRI site of the plasmid pBD64. Subcloning various lengths of the B. licheniformis DNA has localised the gene to a 2550-bp BclI fragment. We present evidence that the cloned fragment codes for a B. licheniformis heat-stable alpha-amylase with a temperature optimum of 93 degrees C. The foreign gene is expressed efficiently in B. subtilis and is stably maintained.
一种为枯草芽孢杆菌开发的宿主-质粒克隆系统已被用于分离携带地衣芽孢杆菌DNA的重组质粒，这些质粒赋予枯草芽孢杆菌α-淀粉酶阴性突变体α-淀粉酶活性。这些质粒在质粒pBD64的EcoRI位点含有一个3550 bp的插入片段。亚克隆地衣芽孢杆菌DNA的不同长度片段已将该基因定位到一个2550 bp的BclI片段上。我们提供的证据表明，克隆片段编码一种最适温度为93℃的地衣芽孢杆菌热稳定α-淀粉酶。该外源基因在枯草芽孢杆菌中高效表达并稳定维持。
