1998
DOI: 10.1016/s0378-1097(97)00500-4
|View full text |Cite
|
Sign up to set email alerts
|

Construction of antibiotic resistance cassettes with multiple paired restriction sites for insertional mutagenesis of Haemophilus influenzae

Abstract: Insertional mutagenesis of cloned genes coupled with site specific recombination into the genome of the parent organism is an ideal method for characterizing gene function. In this paper we describe the production and utility of two antibiotic resistance cassettes for use in Haemophilus influenzae. The mutagenic elements encode resistance to chloramphenicol or spectinomycin. Multiple paired restriction enzyme sites bound both cassettes. Use of these constructs to create mutants in H. influenzae demonstrated th… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
3
1

Citation Types

1
16
0

Year Published

2000
2000
2011
2011

Publication Types

Select...
10

Relationship

0
10

Authors

Journals

citations
Cited by 16 publications
(17 citation statements)
references
References 10 publications
1
16
0
Order By: Relevance
“…In contrast, PCR products of 1.6 kb were generated in isogenic mutants tested with HagF 3760 and HagR 4153 (data not shown). This 1.2-kb difference in the size of amplicons is consistent with the size of the Spec r cassette (56). To confirm that the Spec r cassette is inserted within the hag gene, we used primer sets in which one primer anneals within the hag gene, whereas the other binds to one of the terminal regions of Spec r .…”
Section: Methodssupporting
confidence: 59%
“…In contrast, PCR products of 1.6 kb were generated in isogenic mutants tested with HagF 3760 and HagR 4153 (data not shown). This 1.2-kb difference in the size of amplicons is consistent with the size of the Spec r cassette (56). To confirm that the Spec r cassette is inserted within the hag gene, we used primer sets in which one primer anneals within the hag gene, whereas the other binds to one of the terminal regions of Spec r .…”
Section: Methodssupporting
confidence: 59%
“…Complementation was performed by using a derivative of the shuttle vector pGZRS-39A (46), in which the kanamycin resistance gene was replaced by the spectinomycin resistance gene from pSPECR (48). The pGZRS-39A plasmid backbone, without the kanamycin resistance gene, was amplified by using primers 11 and 12 (Table 1).…”
Section: Methodsmentioning
confidence: 99%
“…After digestion with BamHI, this 2.2-kb DNA fragment was ligated into pBS KS(ϩ). A 0.4-kb BglII fragment was removed from the middle of this partial uspA2 ORF and was replaced with a cartridge encoding resistance to spectinomycin (42), yielding the plasmid pELU2P44SPEC. EcoRI-digested pELU2P44SPEC was used to electroporate the M. catarrhalis uspA1 mutants 012E.1 and TTA37.1.…”
Section: Methodsmentioning
confidence: 99%