1999
DOI: 10.1099/13500872-145-6-1307
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Constructs for insertional mutagenesis, transcriptional signal localization and gene regulation studies in root nodule and other bacteria

Abstract: * Office of theCassettes have been developed that contain an antibiotic resistance marker with and without a promoterless gusA reporter gene. The nptll (encoding kanamycin resistance) or aacCl (encoding gentamicin resistance) genes were equipped with the tac promoter (PtaC) and the trpA terminator ( T , ) and then cloned between Not1 sites to construct the CAS-Nm (Ptac-npt//-TtpA) and CAS-Gm ( Pt , CP, , , c/ -aacC/ -T, )cassettes. The markers were also cloned downstream to a modified promoterless Escherich… Show more

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Cited by 118 publications
(121 citation statements)
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“…A348 contains the same genome components as C58 except for the Ti plasmid, which originated from strain A6. The virB1 promoter was amplified and cloned into the pFUS1 vector to generate a virB1::gusA transcriptional gene fusion (36). Plasmid pWT160 (37) was used to measure virG expression.…”
Section: Methodsmentioning
confidence: 99%
“…A348 contains the same genome components as C58 except for the Ti plasmid, which originated from strain A6. The virB1 promoter was amplified and cloned into the pFUS1 vector to generate a virB1::gusA transcriptional gene fusion (36). Plasmid pWT160 (37) was used to measure virG expression.…”
Section: Methodsmentioning
confidence: 99%
“…Transductions with M12 were performed as described previously (15). Random mini-Tn5-GusNm (mTn5) mutagenesis and triparental matings were performed as described previously (13,37). The protocols of Sambrook and Russell (39) were used for routine manipulations of plasmid and chromosomal DNAs.…”
Section: Methodsmentioning
confidence: 99%
“…Amplification primers were designed containing PstI restriction enzyme recognition sites at the ends, and they were digested with PstI and cloned into pJQ200mp18 (Quandt & Hynes, 1993) digested with PstI, generating pMTA1 and pMTA2 respectively. pCRS487 (Reeve et al, 1999) was digested with NotI, and the gusA-nptII cartridge from this plasmid was cloned directly into the NotI site in pMTA1 or was blunt-ended with Klenow enzyme and cloned into the blunted XhoI site in pMTA2. pMTA5 was generated by digesting pHP45V (Prentki & Krisch, 1984) with SmaI and cloning the spectinomycin resistance cassette into blunted NotI-digested pMTA1.…”
Section: Methodsmentioning
confidence: 99%
“…Plasmid pCRS538 (Reeve et al, 1999) was used to mutagenize strain MTA100. The plasmid was delivered by conjugation from E. coli S17-1, and exconjugants were selected using gentamicin and kanamycin.…”
Section: Methodsmentioning
confidence: 99%