Integration of repeated sequences (pBR322) in the Bacillus subtilis 168 chromosome without affecting the genome structure

Mitsuhiro Itaya

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The Escherichia coli plasmid pBR322 sequence (4363 bp) was integrated at the met, pro, or leuB locus of the Bacillus subtilis chromosome without duplication of the flanking chromosomal regions. The integrated pBR322 was stably maintained as part of the chromosome regardless of its orientation or location. It was found that a DNA segment as large as 17 kb cloned in pBR322 can be readily transferred to the B. subtilis chromosome by transformation. It was demonstrated that a second pBR322 sequence could be effectively introduced at different regions of the chromosome by sequential transformation using chromosomal DNA isolated from a strain that had already acquired a pBR322 sequence at a different locus. Similarly, a third pBR322 sequence could be introduced. By this method, two or three pBR322 sequences can be incorporated at unlinked loci without affecting the overall structure of the B. subtilis genome.

Original languageEnglish
Pages (from-to)287-297
Number of pages11
JournalMGG Molecular & General Genetics
Volume241
Issue number3-4
DOIs
Publication statusPublished - 1993 Nov
Externally publishedYes

Fingerprint

Bacillus subtilis
Chromosomes
Genome
Chromosome Duplication
DNA
Plasmids
Escherichia coli

Keywords

  • Bacillus subtilis chromosome
  • Integration
  • pBR322
  • Transformation

ASJC Scopus subject areas

  • Genetics

Cite this

Integration of repeated sequences (pBR322) in the Bacillus subtilis 168 chromosome without affecting the genome structure. / Itaya, Mitsuhiro.

In: MGG Molecular & General Genetics, Vol. 241, No. 3-4, 11.1993, p. 287-297.

Research output: Contribution to journalArticle

@article{d39bcb14384b43609038e93bdd8f0589,
title = "Integration of repeated sequences (pBR322) in the Bacillus subtilis 168 chromosome without affecting the genome structure",
abstract = "The Escherichia coli plasmid pBR322 sequence (4363 bp) was integrated at the met, pro, or leuB locus of the Bacillus subtilis chromosome without duplication of the flanking chromosomal regions. The integrated pBR322 was stably maintained as part of the chromosome regardless of its orientation or location. It was found that a DNA segment as large as 17 kb cloned in pBR322 can be readily transferred to the B. subtilis chromosome by transformation. It was demonstrated that a second pBR322 sequence could be effectively introduced at different regions of the chromosome by sequential transformation using chromosomal DNA isolated from a strain that had already acquired a pBR322 sequence at a different locus. Similarly, a third pBR322 sequence could be introduced. By this method, two or three pBR322 sequences can be incorporated at unlinked loci without affecting the overall structure of the B. subtilis genome.",
keywords = "Bacillus subtilis chromosome, Integration, pBR322, Transformation",
author = "Mitsuhiro Itaya",
year = "1993",
month = "11",
doi = "10.1007/BF00284680",
language = "English",
volume = "241",
pages = "287--297",
journal = "Molecular Genetics and Genomics",
issn = "1617-4615",
publisher = "Springer Verlag",
number = "3-4",

}

TY - JOUR

T1 - Integration of repeated sequences (pBR322) in the Bacillus subtilis 168 chromosome without affecting the genome structure

AU - Itaya, Mitsuhiro

PY - 1993/11

Y1 - 1993/11

N2 - The Escherichia coli plasmid pBR322 sequence (4363 bp) was integrated at the met, pro, or leuB locus of the Bacillus subtilis chromosome without duplication of the flanking chromosomal regions. The integrated pBR322 was stably maintained as part of the chromosome regardless of its orientation or location. It was found that a DNA segment as large as 17 kb cloned in pBR322 can be readily transferred to the B. subtilis chromosome by transformation. It was demonstrated that a second pBR322 sequence could be effectively introduced at different regions of the chromosome by sequential transformation using chromosomal DNA isolated from a strain that had already acquired a pBR322 sequence at a different locus. Similarly, a third pBR322 sequence could be introduced. By this method, two or three pBR322 sequences can be incorporated at unlinked loci without affecting the overall structure of the B. subtilis genome.

AB - The Escherichia coli plasmid pBR322 sequence (4363 bp) was integrated at the met, pro, or leuB locus of the Bacillus subtilis chromosome without duplication of the flanking chromosomal regions. The integrated pBR322 was stably maintained as part of the chromosome regardless of its orientation or location. It was found that a DNA segment as large as 17 kb cloned in pBR322 can be readily transferred to the B. subtilis chromosome by transformation. It was demonstrated that a second pBR322 sequence could be effectively introduced at different regions of the chromosome by sequential transformation using chromosomal DNA isolated from a strain that had already acquired a pBR322 sequence at a different locus. Similarly, a third pBR322 sequence could be introduced. By this method, two or three pBR322 sequences can be incorporated at unlinked loci without affecting the overall structure of the B. subtilis genome.

KW - Bacillus subtilis chromosome

KW - Integration

KW - pBR322

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=0027486880&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027486880&partnerID=8YFLogxK

U2 - 10.1007/BF00284680

DO - 10.1007/BF00284680

M3 - Article

VL - 241

SP - 287

EP - 297

JO - Molecular Genetics and Genomics

JF - Molecular Genetics and Genomics

SN - 1617-4615

IS - 3-4

ER -