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An improved counterselection cassette for use in Haemophilus influenzae |
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| Authors: | Johnston Jason W |
| Institution: | |
| Abstract: | Counterselectable cassettes are extremely useful in molecular biology and allow for the creation of unmarked deletion mutants or the introduction of point mutations. I have constructed an inducible sacB cassette, using the tetracycline repressor. When used in tandem with a kanamycin-resistance marker, the cassette was successful in creating unmarked mutants in Haemophilus influenzae. The inducible nature of the cassette avoids some of the common problems associated with the utilization of sacB in counterselection. |
| Keywords: | LB Luria-Bertani NTHi nontypeable Haemophilus influenzae NAD nicotinamide adenine dinucleotide sBHI supplemented brain heart Infusion cTc chlortetracycline PCR polymerase chain reaction cAMP adenosine 3&prime 5&prime -cyclic monophosphate DNA deoxyribonucleic acid Rib ribostamycin REN restriction endonuclease bp base pair Suc sucrose SOEing splicing by overlapping extension |
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