Mahler I, Warburg R, Tipper D J, Halvorson H O
J Gen Microbiol. 1984 Feb;130(2):411-21. doi: 10.1099/00221287-130-2-411.
A recombinant cosmid clone was isolated from a library created from cosmid pQB79-1 and Bacillus subtilis DNA, and a 15 kb BamHI fragment derived from the cloned insert was transferred to the vector pHV33. The recombinant clone, pRC12, was capable of complementing eight auxotrophic markers in the spoIIA-tyrA region of the B. subtilis chromosome (map positions 205-210). It also complemented eight of nine markers in the spoIIA locus. The exception, spoIIA176, is the most distal marker from lysine. Although pRC12 failed to complement sporulation defects in spoVA or spoIVA (spoIIA+) strains, subclones of pRC12, lacking a functional spoIIA gene, did complement these mutations. pRC12 inhibited sporulation in a spo+ recE strain, possibly due to the presence of multiple functional spoIIA genes. Both the original cosmid and pRC12 were unstable in Escherichia coli and B. subtilis. Antibiotic selection of the vector resulted in extensive deletion of the insert, while selection for insert function in B. subtilis invariably led to loss of the chloramphenicol resistance vector function.
从由黏粒pQB79-1和枯草芽孢杆菌DNA构建的文库中分离出一个重组黏粒克隆，并将源自克隆插入片段的15 kb BamHI片段转移至载体pHV33。重组克隆pRC12能够互补枯草芽孢杆菌染色体spoIIA-tyrA区域（图谱位置205-210）中的八个营养缺陷型标记。它还能互补spoIIA基因座中九个标记中的八个。例外的是spoIIA176，它是距赖氨酸最远的标记。尽管pRC12不能互补spoVA或spoIVA（spoIIA+）菌株中的芽孢形成缺陷，但缺乏功能性spoIIA基因的pRC12亚克隆确实能互补这些突变。pRC12在spo+ recE菌株中抑制芽孢形成，这可能是由于存在多个功能性spoIIA基因。原始黏粒和pRC12在大肠杆菌和枯草芽孢杆菌中均不稳定。对载体进行抗生素选择会导致插入片段大量缺失，而在枯草芽孢杆菌中选择插入片段功能则总是导致氯霉素抗性载体功能丧失。
