Construction of Escherichia coli Bifidobacterium longum shuttle vector transforming B-longum 105-A and 108-A

Construction of Escherichia coli Bifidobacterium longum shuttle vector transforming B-longum 105-A and 108-A
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DOI:
10.1271/bbb.61.1211
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发表时间:
1997-07-01
影响因子:
1.6
通讯作者:
Kano, Y
Kano, Y
中科院分区:
工程技术4区
文献类型:
--
作者:
Matsumura, H;Takeuchi, A;Kano, Y

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通过将来自粪肠球菌的长双歧杆菌质粒和编码壮观霉素腺苷酸转移酶 AAD(9) 的基因克隆到大肠杆菌载体 pBR322 中,构建了穿梭载体 pBLES100。使用在对数生长期收获的长双歧杆菌 105-A,在 10.0 kV/cm、200 Omega 和 25 mu F 的最佳条件下,以 2.2 x 10(4) 转化子/μ g DNA 或 6.9 x 10(-5) 转化子/细胞/μ g DNA 的效率获得带有该质粒的稳定转化子。
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/mu g DNA or 6.9 x 10(-5) transformants/cell/mu g DNA under the optimal conditions of 10.0 kV/cm, 200 Omega, and 25 mu F, using B. longum 105-A harvested at late log phase of growth.