Efficient cloning and engineering of giant DNAs in a novel Bacillus subtilis genome vector

被引：35

作者：

Itaya, M

Nagata, T

Shiroishi, T

Fujita, K

Tsuge, K

机构：

[1] Mitsubishi Kasei Inst Life Sci, Machida, Tokyo 1948511, Japan

[2] Natl Inst Genet, Mammalian Genet Lab, Mishima, Shizuoka 4118540, Japan

来源：

JOURNAL OF BIOCHEMISTRY | 2000年 / 128卷 / 05期

关键词：

genome vector; positive selection; recombination; transformation;

D O I：

10.1093/oxfordjournals.jbchem.a022825

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The Genome of Bacillus subtilis 168 was used for cloning and engineering of large-sized DNAs, A mouse genomic DNA of approximately 120 kb was clotted into a locus of the B. subtilis genome by ordered assembly of 20- to 50-kb mouse DNA segments. Cloned mouse DNA, maintained stably, was engineered through B, subtilis transformation and recombination, Creation of an I-PpoI recognition sequence at both ends of the insert facilitated its isolation by pulsed field gel electrophoresis, The basic concept of genome vector technology is suited to the handling of DNAs larger than. 100 kb.

引用

页码：869 / 875

页数：7

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