Functional organization of plasmid pKM101

Langer, Pamela J.; Shanabruch, William G.; Walker, Graham C.

doi:10.1128/jb.145.3.1310-1316.1981

Cited by 95 publications

(34 citation statements)

References 26 publications

(40 reference statements)

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“…Plamd constructions. Plasmid pSE200 was constructed by cloning BglII-digested pGW270 (15) DNA into BamHI-cleaved pMC874 and transforming into GW1000, selecting for kanamycin resistance (Kmr) on plates containing Xgal. Kmr blue colonies were screened for ampicilhin sensitivity (Aps) to eliminate derivatives containing the BgIII fragment of pGW270 carrying the bla gene, which codes for 1-lactamase.…”

Section: Methodsmentioning

confidence: 99%

The muc genes of pKM101 are induced by DNA damage

Elledge¹,

Walker²

1983

J Bacteriol

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A gene fusion was constructed in vitro that resulted in the synthesis of a hybrid protein consisting of the amino-terminal segment of the MucB protein of the mutagenesis-enhancing plasmid pKM101 joined to an enzymatically active carboxy-terminal segment of the beta-galactosidase protein. In strains bearing this fusion, beta-galactosidase activity was induced by UV radiation and other DNA-damaging agents. A genetic analysis of the regulation of expression of the phi (mucB'-lacZ') fusion was consistent with the LexA protein acting as the direct repressor of the mucB gene. Examination of the expression of the mucA and phi (mucB'-lacZ') gene products in maxicells in the presence and absence of a high-copy-number plasmid carrying the lexA+ gene demonstrated that lexA regulated both the mucA and mucB genes, thus supporting our conclusion that the two genes are organized in an operon with the mucA gene transcribed first. An analysis of the effects of the recA430(lexB30) mutation on muc expression led to the discovery of the differential ability of the recA430 gene product to induce expression of a dinB::Mu d1(Ap lac) fusion located on the chromosome and the same phi (dinB'-lacZ+) fusion cloned into plasmid pBR322. Models to account for the role of the recA430 allele on the expression of damage-inducible genes and on mutagenesis are discussed.

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Section: Methodsmentioning

confidence: 99%

The muc genes of pKM101 are induced by DNA damage

Elledge¹,

Walker²

1983

J Bacteriol

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“…Plasmid pTD59 (10 ng/#l) was irradiated with 225 ergs/mm e or 450 ergs/mm 2 of 254 um light using four 15W germicidal light bulbs (GI5TS; General Electric Co., Wilmington, MA) and then placed on ice. E. coli strain AB1886 (pGNr (Gimble and Saner, 1985;Langer et al, 1981) was grown in 10 mi LB containing 30 #g/mi kanamycin to 100 Klett units (approximately 5 • l0 s cells/ml). Cells were pelleted at 6,000 rpm for 10 rain in an SS-34 rotor (Sorvall Instruments, Wilmington, DE), washed in )~dll, pelleted again and resospended in 5 mi Xdil.…”

Section: Mutagenesismentioning

confidence: 99%

“…The E. coli strain is deficient in excision repair (uvrA-) and thus mutations were introduced during the error-prone repair of the damaged DNA. The efficiency of mutagenesis was enhanced by the mutator plasmid pGW249 (Gimble and Sauer, 1985;Langer et al, 1981), which is a derivative of plasmid pKM101 (Walker, 1977). Plasmid pKM101 enhances mutagenesis in E. coli 3-10-fold (Glickman, 1983).…”

Section: Isolation Of a Temperature-sensitive Mutation In Cmd1mentioning

confidence: 99%

A temperature-sensitive calmodulin mutant loses viability during mitosis

Davis

1992

Trends in Cell Biology

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“…The R-factor strains (TA 97, TA 98, TA 100 and TA 102) should be tested routinely for the presence of the ampicillin resistance factor because the plasmid is somewhat unstable and can be lost from the bacteria [15]. Specific regions of the pKMlOl DNA that are essential for enhancement of UV and chemical mutagenesis, replication, and ampicillin resistance have been identified [16]. 0,1 ml Samples that were got from night cultures of test strains spread to plaques with nutrient agar for control of RFA mutation.…”

Section: Control Of Genetic Specialities Of Strainsmentioning

confidence: 99%

Determination of Genotoxic Pollution of Some Hospital Wastewater with Salmonella Ames Test

Atasoy¹,

Karakeçe²,

Petek³

et al. 2012

JWARP

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Wastewater of hospitals contains materials that would be a threat to alive. These water needs to be checked by a biological purification before leaving to nature. Hospital wastewater has differences than domestic waste because of especially blood, body waste, drugs, chemicals, medical device waste and radioactive materials. We aimed to determine genotoxic effects of total pollution in hospital wastewater on alive by Salmonella microsome test method. In this study, we decided on three hospitals which weren’t checked as biological purification of waste. The samples were taken for six 1-week periods between March 2009 and June 2009. Mutagenite studies of samples taken from hospitals were made with ,<i>Salmonella typhimurium</i> TA 98 and TA 100. Wastewater samples were evaporated. 27 different test materials were prepared using DMSO, ethanol and acetone solvents, two different MGA plaques were used for each test material. Each experiment was made for 3 times with known results of mutagens and we made it ready for “Ames” test method. We had genotoxicity 50% in Istanbul University Medical Faculty Hospital, 56% in Haseki Hospital and 61% in Vak?f Gureba Hospital. According to three hospitals result there are 9 positives, 9 negatives in DMSO; 9 positives, 9 negatives in ethanol; 12 positives, 6 negatives in acetone. These values are totally 56%. Our results give important information about mutagenic effect of total pollution in hospital wastewater. It is first time researched in Turkey that effect on DNA of pollution is from hospital wastewaters. In prospective studies, it is necessary to use this system as a method to monitor mutagenic genotoxic pollution in hospital wastewaters. These kinds of studies present applicability and importance of our method because of placing in the literature. Method constitutes a new approach to check mutagenite of pollution in hospital wastewater

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Functional organization of plasmid pKM101

Cited by 95 publications

References 26 publications

The muc genes of pKM101 are induced by DNA damage

The muc genes of pKM101 are induced by DNA damage

A temperature-sensitive calmodulin mutant loses viability during mitosis

Determination of Genotoxic Pollution of Some Hospital Wastewater with Salmonella Ames Test

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