In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries

W. R. Jacobs, J. F. Barrett, J. E. Clark-Curtiss, R. Curtiss

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Strains of Escherichia coli K-12 were constructed that permitted the amplification of in vitro-packaged recombinant cosmid-transducing particles by in vivo repackaging of recombinant cosmid molecules. Thermal induction of these thermoinducible, excision-defective lysogens containing recombinant cosmid molecules yielded high titers of packaged recombinant cosmids and low levels of PFU. These strains were used to amplify packaged recombinant cosmid libraries of Mycobacterium leprae, Mycobacterium vaccae, Salmonella typhimurium, and Streptococcus mutans DNA. Contiguous and noncontiguous libraries were compared for the successful identification of cloned genes. Construction of noncontiguous libraries allowed the dissociation of desired genes from genes that were deleterious to the survival of a cosmid recombinant and permitted selection for unliked traits that resulted in a selected phenotype. In vivo repackaging of recombinant cosmids permitted (i) amplification of the original in vitro-packaged collection of transducing particles, (ii) storage of cosmid libraries as phage lysates, (iii) facilitation of complementation screening, (iv) expression analysis of repackaged recombinant cosmids after UV-irradiated cells were infected, (v) in situ enzyme or immunological screening, and (vi) facilitation of recovery of recombinant cosmid molecules containing transposon inserts.

Original languageEnglish (US)
Pages (from-to)101-109
Number of pages9
JournalInfection and Immunity
Volume52
Issue number1
StatePublished - 1986
Externally publishedYes

Fingerprint

Cosmids
Streptococcus mutans
Genomic Library
Salmonella typhimurium
Libraries
Genes
Mycobacterium leprae
Mycobacterium
Bacteriophages
Hot Temperature
Escherichia coli
Phenotype

ASJC Scopus subject areas

  • Immunology

Cite this

In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries. / Jacobs, W. R.; Barrett, J. F.; Clark-Curtiss, J. E.; Curtiss, R.

In: Infection and Immunity, Vol. 52, No. 1, 1986, p. 101-109.

Research output: Contribution to journalArticle

@article{c3de108964694078b77443ae9673a43e,
title = "In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries",
abstract = "Strains of Escherichia coli K-12 were constructed that permitted the amplification of in vitro-packaged recombinant cosmid-transducing particles by in vivo repackaging of recombinant cosmid molecules. Thermal induction of these thermoinducible, excision-defective lysogens containing recombinant cosmid molecules yielded high titers of packaged recombinant cosmids and low levels of PFU. These strains were used to amplify packaged recombinant cosmid libraries of Mycobacterium leprae, Mycobacterium vaccae, Salmonella typhimurium, and Streptococcus mutans DNA. Contiguous and noncontiguous libraries were compared for the successful identification of cloned genes. Construction of noncontiguous libraries allowed the dissociation of desired genes from genes that were deleterious to the survival of a cosmid recombinant and permitted selection for unliked traits that resulted in a selected phenotype. In vivo repackaging of recombinant cosmids permitted (i) amplification of the original in vitro-packaged collection of transducing particles, (ii) storage of cosmid libraries as phage lysates, (iii) facilitation of complementation screening, (iv) expression analysis of repackaged recombinant cosmids after UV-irradiated cells were infected, (v) in situ enzyme or immunological screening, and (vi) facilitation of recovery of recombinant cosmid molecules containing transposon inserts.",
author = "Jacobs, {W. R.} and Barrett, {J. F.} and Clark-Curtiss, {J. E.} and R. Curtiss",
year = "1986",
language = "English (US)",
volume = "52",
pages = "101--109",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "1",

}

TY - JOUR

T1 - In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries

AU - Jacobs, W. R.

AU - Barrett, J. F.

AU - Clark-Curtiss, J. E.

AU - Curtiss, R.

PY - 1986

Y1 - 1986

N2 - Strains of Escherichia coli K-12 were constructed that permitted the amplification of in vitro-packaged recombinant cosmid-transducing particles by in vivo repackaging of recombinant cosmid molecules. Thermal induction of these thermoinducible, excision-defective lysogens containing recombinant cosmid molecules yielded high titers of packaged recombinant cosmids and low levels of PFU. These strains were used to amplify packaged recombinant cosmid libraries of Mycobacterium leprae, Mycobacterium vaccae, Salmonella typhimurium, and Streptococcus mutans DNA. Contiguous and noncontiguous libraries were compared for the successful identification of cloned genes. Construction of noncontiguous libraries allowed the dissociation of desired genes from genes that were deleterious to the survival of a cosmid recombinant and permitted selection for unliked traits that resulted in a selected phenotype. In vivo repackaging of recombinant cosmids permitted (i) amplification of the original in vitro-packaged collection of transducing particles, (ii) storage of cosmid libraries as phage lysates, (iii) facilitation of complementation screening, (iv) expression analysis of repackaged recombinant cosmids after UV-irradiated cells were infected, (v) in situ enzyme or immunological screening, and (vi) facilitation of recovery of recombinant cosmid molecules containing transposon inserts.

AB - Strains of Escherichia coli K-12 were constructed that permitted the amplification of in vitro-packaged recombinant cosmid-transducing particles by in vivo repackaging of recombinant cosmid molecules. Thermal induction of these thermoinducible, excision-defective lysogens containing recombinant cosmid molecules yielded high titers of packaged recombinant cosmids and low levels of PFU. These strains were used to amplify packaged recombinant cosmid libraries of Mycobacterium leprae, Mycobacterium vaccae, Salmonella typhimurium, and Streptococcus mutans DNA. Contiguous and noncontiguous libraries were compared for the successful identification of cloned genes. Construction of noncontiguous libraries allowed the dissociation of desired genes from genes that were deleterious to the survival of a cosmid recombinant and permitted selection for unliked traits that resulted in a selected phenotype. In vivo repackaging of recombinant cosmids permitted (i) amplification of the original in vitro-packaged collection of transducing particles, (ii) storage of cosmid libraries as phage lysates, (iii) facilitation of complementation screening, (iv) expression analysis of repackaged recombinant cosmids after UV-irradiated cells were infected, (v) in situ enzyme or immunological screening, and (vi) facilitation of recovery of recombinant cosmid molecules containing transposon inserts.

UR - http://www.scopus.com/inward/record.url?scp=0022637815&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022637815&partnerID=8YFLogxK

M3 - Article

VL - 52

SP - 101

EP - 109

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 1

ER -