Addition, deletion, and substitution of long nonhomologous deoxyribonucleic acid segments by genetic transformation of Haemophilus influenzae.

Journal of Bacteriology
J H Stuy, R B Walter

Abstract

A complete EcoRI digest of Haemophilus influenzae phage HP1 deoxyribonucleic acid (DNA) was mixed with incomplete digests of various H. influenzae R plasmids, sealed with T4 ligase, and transformed into an HP1 lysogen. Most of the chloramphenicol- and tetracycline-resistant transformants did not produce phage although they possessed all the phage genes examined. They also did not transfer antibiotic resistance by conjugation. DNA lysates from them transformed other lysogens to resistance and to loss of phage production at different but quite high frequencies (addition of long DNA segments). They themselves could be transformed efficiently to strains with a wild prophage (deletion of long DNA segments). It was concluded that lysogenic cultures had been constructed with various DNA inserts in their prophages carrying antibiotic resistance genes from the R plasmids. The site of insertion was determined by genetic crosses. DNAs with inserts that transferred with lower efficiency were more sensitive to ultraviolet radiation. This supports the view that insert transfer efficiencies reflect the sizes of the insert.

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Citations

Apr 1, 1983·Photochemistry and Photobiology·J H Stuy, R B Walter
Aug 11, 2011·PLoS Pathogens·Joshua Chang MellRosemary J Redfield
May 1, 1986·Molecular & General Genetics : MGG·J H Stuy, R B Walter
Sep 8, 1998·FEMS Microbiology Reviews·K M NielsenJ D van Elsas
Feb 27, 2014·G3 : Genes - Genomes - Genetics·Joshua Chang MellRosemary J Redfield
Jun 1, 1987·Journal of Bacteriology·G L WilliamsD McCarthy
Apr 1, 1986·Journal of Bacteriology·J H Stuy

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