Cotransformation of linear chromosomal DNA and plasmid DNA in Escherichia coli

Bergmans, Hans

doi:10.1016/0378-1097(80)90126-3

Cited by 3 publications

(4 citation statements)

References 7 publications

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“…The amplicon was PCR amplified from total DNA obtained from A. rhizogenes A4, purified using QIAquick PCR Purification kit (QIAGEN) and digested with restriction enzymes ( SmaI , SalI , Roche) for subsequent ligation (T4 DNA Ligase, Thermo Scientific) into pBluscript II‐SK+. Competent E. coli DH5α cells were transformed with the ligation mixture using heat shock method (Bergmans et al, 1980). The competent cells of E. coli DH5α were prepared following the method of Green and Rogers (2013), with an addition of 10 mM (3‐(N‐Morpholino)‐propanesulfonic acid [MOPS]) to the transformation buffer‐I.…”

Section: Methodsmentioning

confidence: 99%

“…The cloned rolB was excised from rolB ‐pBluescript II SK+ recombinant vector by restriction enzyme digestion ( Sma I, Sal I; Roche) and introduced in a modified pCAMBIA1301 (MpCAMBIA1301) (Mukherjee et al, 2019) plant transformation vector by ligation. Subsequent transformation into competent E. coli DH5α cells using heat shock method (Bergmans et al, 1980) was performed. Positive recombinants were selected by growth on LB‐Kanamycin (50 mg L −1 ) plates and screened by colony PCR using rolB specific primers.…”

Section: Methodsmentioning

confidence: 99%

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The rolB‐transgenic Nicotiana tabacum plants exhibit upregulated ARF7 and ARF19 gene expression

et al. 2022

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Agrobacterium rhizogenes root oncogenic locus B (rolB) is known to induce hairy roots along with triggering several physiological and morphological changes when present as a transgene. However, it is still unknown how this gene triggers these changes within the plant system. In this study, the effect of rolB in-planta, when present as a transgene, was assessed on the gene expression levels of auxin response factors (ARFs)-transcription factors which are key players in auxin-mediated responses. The goal was to uncover Auxin/ARF-driven transcriptional networks potentially active and working selectively, if any, in rolB transgenic background, which might potentially be associated with hairy root development. Hence, the approach involved establishing rolB-transgenic Nicotiana tabacum plants, selecting ARFs (NtARFs) for context-relevance using bioinformatics followed by gene expression profiling. It was observed that out of the chosen NtARFs, NtARF7 and NtARF19 exhibited a consistent pattern of gene upregulation across organ types. In order to understand the significance of these selective gene upregulation, ontology-based transcriptional network maps of the differentially and nondifferentially expressed ARFs were constructed, guided by co-expression databases. The network maps suggested that NtARF7-NtARF19 might have major deterministic, underappreciated roles to play in root development in a rolB-transgenic background-as observed by higher number of "root-related" biological processes present as nodes compared to network maps for similarly constructed other non-differentially expressed ARFs. Based on the inferences drawn, it is hypothesized that rolB, when present as a transgene, might drive hairy root development by selective induction of NtARF7 and NtARF19, suggesting a functional link between the two, leading to the specialized and characteristic rolBassociated traits.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The rolB‐transgenic Nicotiana tabacum plants exhibit upregulated ARF7 and ARF19 gene expression

et al. 2022

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“…Under optimal conditions, with strains with a high efficiency of transformation, uptake of plasmid DNA was seen only in about 0.1 to 1% of the cells. The same individual cells preferentially took up a second plasmid (1,13). The amount of DNA introduced into the periplasm was much higher than that found in the cytoplasm (29).…”

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confidence: 92%

Ca2+-induced permeabilization of the Escherichia coli outer membrane: comparison of transformation and reconstitution of binding-protein-dependent transport

Bukau¹,

Brass²,

Boos³

1985

J Bacteriol

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Ca2+ treatment renders the outer membrane of Escherichia coli reversibly permeable for macromolecules. We investigated whether Ca2+-induced uptake of exogenous protein into the periplasm occurs by mechanisms similar to Ca2+-induced uptake of DNA into the cytoplasm during transformation. Protein import through the outer membrane was monitored by measuring reconstitution of maltose transport after the addition of shock fluid containing maltose-binding protein. DNA import through the outer and inner membrane was measured by determining the efficiency of transformation with plasmid DNA. Both processes were stimulated by Preparation of shock fluid. Shock fluid of maltose-induced cells of strain poplO80 [HfrG6 lamB102(Am) metA trpE(Am) galE galY] (18), containing between 30 and 50% of total protein as MBP, was prepared by osmotic shock by the 61 on August 5, 2020 by guest

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“…Competence for DNA uptake in Escherichia coli transformation is usually achieved by a heat shock from 0°C to temperatures between 30°C and 42°C, in the presence of Ca2+-ions [1][2][3][4][5][6][7][8]. Under optimal conditions, only a small proportion (0.1-1%) of the cell population becomes competent [6,9]. The growth phase of batch cultures as well as the growth rate and nutrient limitation in chemostat cultures greatly influence competence for plasmid transformation [10,11].…”

Section: Introductionmentioning

confidence: 99%

The influence of phase transition of membrane lipids on uptake of plasmid DNA in transformation

Vandie¹,

Vanoosterhout²,

Bergmans³

et al. 1983

FEMS Microbiology Letters

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Cotransformation of linear chromosomal DNA and plasmid DNA in Escherichia coli

Cited by 3 publications

References 7 publications

The rolB‐transgenic Nicotiana tabacum plants exhibit upregulated ARF7 and ARF19 gene expression

The rolB‐transgenic Nicotiana tabacum plants exhibit upregulated ARF7 and ARF19 gene expression

Ca2+-induced permeabilization of the Escherichia coli outer membrane: comparison of transformation and reconstitution of binding-protein-dependent transport

The influence of phase transition of membrane lipids on uptake of plasmid DNA in transformation

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