Davison J, Heusterspreute M, Chevalier N, Ha-Thi V, Brunel F
Gene. 1987;51(2-3):275-80. doi: 10.1016/0378-1119(87)90316-7.
The construction of a new wide-host-range, restriction-site bank, cosmid-cloning vehicle (pJRD215) is described. The wide-host-range properties and the ability to be transferred by conjugation, extend genetic engineering to those Gram-negative species that cannot be transformed. The vector permits the cloning of genes from Gram-negative bacteria using a complementation screening procedure in a mutant host. This procedure is simplified by the possibility of construction of a cosmid gene bank so that only a few hundred clones need to be screened. Subsequent subcloning of the gene of interest is facilitated by the presence of at least 23 unique cloning sites.
本文描述了一种新型广宿主范围、限制酶切位点文库黏粒克隆载体（pJRD215）的构建。其广宿主范围特性以及通过接合转移的能力，将基因工程扩展到了那些无法进行转化的革兰氏阴性菌。该载体允许使用突变宿主中的互补筛选程序从革兰氏阴性细菌中克隆基因。通过构建黏粒基因文库的可能性简化了这一程序，使得只需筛选几百个克隆。至少23个独特克隆位点的存在便于后续对感兴趣基因的亚克隆。
