Conjugal transfer replication of R64drd11 plasmid DNA in the donor cells of Escherichia coli K-12

1982 ◽  
Vol 187 (2) ◽  
pp. 305-309 ◽  
Author(s):  
Elaine J. Davis ◽  
Jacquelyn Henry
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna ◽  
Conjugal Transfer ◽  
Donor Cells ◽  
K 12

scholarly journals The Conjugation Window in an Escherichia coli K-12 Strain with an IncFII Plasmid

2020 ◽  
Vol 86 (17) ◽  
Author(s):  
Brendan Headd ◽  
Scott A. Bradford
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antibiotic Resistance Genes ◽  
Time Frame ◽  
Environmental Aspect ◽  
Content Type ◽  
E Coli ◽  
Donor Cells ◽  
K 12 ◽  
Regulatory Controls

ABSTRACT Many studies have examined the role that conjugation plays in disseminating antibiotic resistance genes in bacteria. However, relatively little research has quantitively examined and modeled the dynamics of conjugation under growing and nongrowing conditions beyond a couple of hours. We therefore examined growing and nongrowing cultures of Escherichia coli over a 24-h period to understand the dynamics of bacterial conjugation in the presence and absence of antibiotics with pUUH239.2, an IncFII plasmid containing multiantibiotic- and metal-resistant genes. Our data indicate that conjugation occurs after E. coli cells divide and before they have transitioned to a nongrowing phase. The result is that there is only a small window of opportunity for E. coli to conjugate with pUUH239.2 under both growing and nongrowing conditions. Only a very small percentage of the donor cells likely are capable of even undergoing conjugation, and not all transconjugants can become donor cells due to molecular regulatory controls and not being in the correct growth phase. Once a growing culture enters stationary phase, the number of capable donor cells decreases rapidly and conjugation slows to produce a plateau. Published models did not provide accurate descriptions of conjugation under nongrowing conditions. We present here a modified modeling approach that accurately describes observed conjugation behavior under growing and nongrowing conditions. IMPORTANCE There has been growing interest in horizontal gene transfer of antibiotic resistance plasmids as the antibiotic resistance crisis has worsened over the years. Most studies examining conjugation of bacterial plasmids focus on growing cultures of bacteria for short periods, but in the environment, most bacteria grow episodically and at much lower rates than in the laboratory. We examined conjugation of an IncFII antibiotic resistance plasmid in E. coli under growing and nongrowing conditions to understand the dynamics of conjugation under which the plasmid is transferred. We found that conjugation occurs in a narrow time frame when E. coli is transitioning from a growing to nongrowing phase and that the conjugation plateau develops because of a lack of capable donor cells in growing cultures. From an environmental aspect, our results suggest that episodic growth in nutrient-depleted environments could result in more conjugation than sustained growth in a nutrient rich environment.

Genomic replacement in Escherichia coli K-12 using covalently closed circular plasmid DNA

Gene ◽  
1990 ◽  
Vol 96 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Kristine L. Oden ◽  
Linda C. DeVeaux ◽  
Cecile Rose T. Vibat ◽  
John E. Croman ◽  
Robert B. Gennis
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna ◽  
Circular Plasmid ◽  
K 12

scholarly journals TEMPERATURE-SENSITIVE MUTANTS FOR THE REPLICATION OF PLASMIDS IN ESCHERICHIA COLI II. PROPERTIES OF HOST AND PLASMID MUTATIONS

Genetics ◽  
1973 ◽  
Vol 74 (1) ◽  
pp. 1-16
Author(s):  
David T Kingsbury ◽  
Donna G Sieckmann ◽  
Donald R Helinski
Keyword(s):  
Escherichia Coli ◽  
Dna Replication ◽  
Cell Growth ◽  
Dna Synthesis ◽  
Plasmid Dna ◽  
Recipient Cell ◽  
Conjugal Transfer ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Group I

ABSTRACT Host mutations in Escherichia coli K12 selected for the temperature-sensitive replication of the bacterial plasmid colicinogenic factor E1 (ColE1) exhibit a pleiotropic effect with respect to the effect of the mutation on other extrachromosomal elements. The mutations also vary with respect to the time of incubation of the cells at 43°C required for complete cessation of COlE1, DNA synthesis. While the synthesis of the bacterial chromosome appears unaffected, supercoiled ColE1 DNA replication stops immediately in some mutants and gradually decreases during several generations of cell growth before stopping in others. Mutations isolated in the ColE1 plasmid resulted in only a gradual cessation of ColE1 DNA synthesis over several generations of cell growth at 43°C. Conjugal transfer of the ColE1 and COlV factors occurs normally in the host mutants when the transfer is carried out at the permissive temperature; however, the presence of a group I mutation in the donor cell prohibited conjugal transfer of either plasmid DNA at 43°C to a normal recipient cell. Similarly, the presence of this mutation in the recipient prevented the establishment of COlE1 or COlV in the mutant recipient cell upon conjugation with a normal donor at 43°C. Various host COlE1, replication mutants carrying either ColE1 or ColE2 were also defective in the mitomycin Cinduced production of colicin E1 or colicin E2 at 43°C. The majority of the host mutations examined exhibited a temperature sensitivity to growth in deoxycholate in addition to the inhibition of plasmid DNA replication, suggesting a membrane alteration in these mutants when grown at the restrictive temperature.

scholarly journals Mechanisms of Strand Replacement Synthesis for Plasmid DNA Transferred by Conjugation

2005 ◽  
Vol 187 (10) ◽  
pp. 3400-3406 ◽  
Author(s):  
Christopher Parker ◽  
Richard Meyer
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna ◽  
Salmonella Enterica ◽  
Single Strand ◽  
Complementary Strand ◽  
E Coli ◽  
Dna Primase ◽  
Donor Cells ◽  
Serovar Typhimurium ◽  
Derivatives Of

ABSTRACT A single strand of plasmid DNA is transferred during conjugation. We examined the mechanism of complementary strand synthesis in recipient cells following conjugative mobilization of derivatives of the IncQ plasmid R1162. A system for electroporation of donor cells, followed by immediate mating, was used to eliminate plasmid-specific replicative functions. Under these conditions, Escherichia coli recipients provided a robust mechanism for initiation of complementary strand synthesis on transferred DNA. In contrast, plasmid functions were important for efficient strand replacement in recipient cells of Salmonella enterica serovar Typhimurium. The mobilizing vector for R1162 transfer, the IncP1 plasmid R751, encodes a DNA primase with low specificity for initiation. This protein increased the frequency of transfer of R751 into Salmonella, but despite its low specificity, it was inactive on the R1162 derivatives. The R751 primase was slightly inhibitory for the transfer of both R751 and R1162 into E. coli. The results show that there is a chromosomally encoded mechanism for complementary strand synthesis of incoming transferred DNA in E. coli, while plasmid-specific mechanisms for this synthesis are important in Salmonella.

Intergeneric conjugal transfer of plasmid DNA from Escherichia coli to Kitasatospora setae , a bafilomycin B 1 producer

2004 ◽  
Vol 181 (4) ◽  
pp. 294-298 ◽  
Author(s):  
Sun-Uk Choi ◽  
Chang-Kwon Lee ◽  
Yong-Il Hwang ◽  
Hiroshi Kinoshita ◽  
Takuya Nihira
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna ◽  
Conjugal Transfer
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