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Direct selection for P1-sensitive mutants of enteric bacteria.

R B Goldberg, R A Bender, S L Streicher

Journal of Bacteriology

|June 1, 1974

Summary

Researchers developed a new method to isolate coliphage P1-sensitive mutants in bacteria like Klebsiella. This technique enhances genetic studies and intergeneric hybrid construction for previously difficult-to-study bacterial genera.

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Area of Science:

Microbiology
Bacteriology
Molecular Biology

Background:

Coliphage P1 is a well-characterized bacteriophage used in bacterial genetics.
Many bacterial genera, including Klebsiella, Enterobacter, Citrobacter, and Erwinia, are naturally resistant to coliphage P1.
Limited genetic manipulation systems exist for these naturally resistant bacterial genera.

Purpose of the Study:

To develop a novel method for isolating coliphage P1-sensitive mutants in bacterial genera typically resistant to this phage.
To enable enhanced genetic studies and manipulation in a broader range of bacterial species.
To expand the possibilities for constructing and studying intergeneric hybrids.

Main Methods:

Utilized the P1clr100KM bacteriophage, which is heat-inducible and confers kanamycin resistance.
Applied a method to select for bacterial mutants exhibiting sensitivity to coliphage P1 infection.
Screened various bacterial genera, including Klebsiella, Enterobacter, Citrobacter, and Erwinia, for P1-sensitive mutants.

Main Results:

Successfully isolated coliphage P1-sensitive mutants in Klebsiella, Enterobacter, Citrobacter, and Erwinia.
Demonstrated the effectiveness of the P1clr100KM phage system for inducing sensitivity.
Established a viable genetic system for previously recalcitrant bacterial genera.

Conclusions:

The developed method significantly expands the utility of coliphage P1 as a genetic tool.
This technique facilitates genetic research in bacterial genera lacking robust manipulation systems.
The study broadens the scope for creating and analyzing intergeneric bacterial hybrids.

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