Charakterisierung eines CaCl2-abhängigen Transfektionssystems vonEscherichia coli und Lambda-Phagen-DNS

Andree, H; Dh, Liebscher

doi:10.1002/jobm.3630180102

Cited by 6 publications

(1 citation statement)

References 24 publications

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“…In contrast, transfection ofE. coli by ~bA or ~ DNA has been reported to be more efficient with late exponential or early stationary phase cultures [ 10,11 ]. At no stage during growth can E. coli cells be transformed without CaC12-treatment.…”

Section: Discussionmentioning

confidence: 99%

Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth

Brown

1979

FEMS Microbiology Letters

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Section: Discussionmentioning

confidence: 99%

Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth

Brown

1979

FEMS Microbiology Letters

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Factors which influence CaCl₂ dependent transfection of Lambda DNA in Escherichia coli K12 recipients

Pfeifer¹,

Pöhlmann²,

Betcke³

et al. 1980

J of Basic Microbiology

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This paper presents further parameters influencing the competence, the process of DNA uptake and the efficiency of plating of CaCI,-treated E. coli K12 strains. We have found that the process of DNA uptake depends not only on the treatment of bacteria with a certain CaClponcentrotion but is also influenced considerably by a shift-down of the CaC1,-concentration in the reaction mixture. The pH of the growth media and of the reaction mixture plays an important role in maintaining of optimal transfection. The efficiency of plating is influenced by the thickness of the top layer and the concentration of bacteria on the plate.Without genetic variation of the strains, by only varying the mentioned factors we could improve the efficiency of CaCl, transfection a t about two orders of magnitude to a maximum of 6 x lo6 pfu/pg DNA. 520-525. YAMAMOTO, K. R., ALBERTS, B. A., BENZINGER, R., LAWHORNE, L. and TREIBER, G., 1970. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large scale virus purification. Virology, 40, 734-744.

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Charakterisierung eines CaCl₂‐abhängigen Transfektionssystems von Escherichia coli und T3‐Phagen‐DNS

Stompe

Michel

Mann

et al. 1982

J of Basic Microbiology

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Transfection by DKA isolated from bacteriophage T3 was studied using Escherichia coli 921/0 as host. The following conditions were found optimal:Competent E . coli 921/0 were obtained by harvesting the bacteria a t the onset of late exponential growth ( 5 x lo8 cells/ml) and treating the latter with 0 . 0 5~ CaCI,. Hereafter, the microbes were suspended in 50 mM Tris-HCI buffer (pH 7.2) and the concentration adjusted to 7 x lo9 cells/mI. T 3 DNA was added and the suspension kept at 0 "C for 15 min. Determination of the number of infectious centers was then carried out in the usual way. The efficiency of transfection under these conditions amounted to lo4 p. f. u./Fg DNA.Preincubation of competent bacteria with T4 DNA a t 0 "C before the addition of T3 DNA reduced the number of infectious centers. However, if T3-and T4 DKA were added simultaneously no decrease of the transfection efficiency occurred. Calf thymus DNA was without influence on transfection.

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Charakterisierung eines CaCl2-abhängigen Transfektionssystems vonEscherichia coli und Lambda-Phagen-DNS

Cited by 6 publications

References 24 publications

Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth

Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth

Factors which influence CaCl₂ dependent transfection of Lambda DNA in Escherichia coli K12 recipients

Charakterisierung eines CaCl₂‐abhängigen Transfektionssystems von Escherichia coli und T3‐Phagen‐DNS

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Charakterisierung eines CaCl2-abhängigen Transfektionssystems vonEscherichia coli und Lambda-Phagen-DNS

Cited by 6 publications

References 24 publications

Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth

Transformation of Escherichia coli C600 by plasmid DNA at different phases of growth

Factors which influence CaCl2 dependent transfection of Lambda DNA in Escherichia coli K12 recipients

Charakterisierung eines CaCl2‐abhängigen Transfektionssystems von Escherichia coli und T3‐Phagen‐DNS

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Factors which influence CaCl₂ dependent transfection of Lambda DNA in Escherichia coli K12 recipients

Charakterisierung eines CaCl₂‐abhängigen Transfektionssystems von Escherichia coli und T3‐Phagen‐DNS