Matsumura H, Takeuchi A, Kano Y
Department of Molecular Genetics, Kyoto Pharmaceutical University, Japan.
Biosci Biotechnol Biochem. 1997 Jul;61(7):1211-2. doi: 10.1271/bbb.61.1211.
A shuttle vector, pBLES100, was constructed by cloning a Bifidobacterium longum plasmid and a gene encoding spectinomycin adenyltransferase AAD(9) from Enterococcus faecalis into the Escherichia coli vector pBR322. Stable transformants with this plasmid were obtained with an efficiency of 2.2 x 10(4) transformants/microgram DNA or 6.9 x 10(-5) transformants/cell/microgram DNA under the optimal conditions of 10.0 kV/cm, 200 omega, and 25 microF, using B. longum 105-A harvested at late log phase of growth.
通过将长双歧杆菌质粒和编码来自粪肠球菌的壮观霉素腺苷转移酶AAD(9)的基因克隆到大肠杆菌载体pBR322中，构建了穿梭载体pBLES100。在10.0 kV/cm、200Ω和25μF的最佳条件下，使用处于生长对数后期收获的长双歧杆菌105-A，以2.2×10⁴转化子/微克DNA或6.9×10⁻⁵转化子/细胞/微克DNA的效率获得了携带该质粒的稳定转化体。
