ISSN:
1617-4623
Keywords:
Key words DNA methyltransferase
;
Restrictionendonuclease
;
Neisseria gonorrhoeae
;
Transformation
;
Mutagenesis
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract A technique that allows for easy identification of transformants of Neisseria gonorrhoeae in the absence of selective pressure has been developed. A suicide vector that contains a gonococcal DNA uptake sequence was constructed to aid in DNA uptake. In this transformation procedure, a limiting number of cells is incubated with an excess amount of DNA, and the mixture is plated onto a non-selective medium. At least 20% of the resulting colonies contained cells that had been transformed. This strategy was utilized to construct specific deletions of the S.NgoI, II, IV, V and VII restriction-modification (R/M) genes. All five deletions were successfully incorporated into the chromosome of FA19, producing strain JUG029. Strain JUG029 could be transformed with non-methylated plasmid DNA while strain FA19 could not be transformed with such DNA. The development of a simple, non-selective transformation technique, coupled with the construction of a strain that is more permissive for DNA-mediated transformation, will aid in genetic manipulations of the gonococcus.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02173639
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