Efficient transformation of Agrobacterium tumefaciens by electroporation

doi:10.1016/0378-1119(90)90452-W

Gene

Volume 90, Issue 1, 31 May 1990, Pages 149-151

https://doi.org/10.1016/0378-1119(90)90452-W Get rights and content

Abstract

High-voltage electroporation was used to transform Agrobacterium tumefaciens strains A136 and A348, reaching the efficiency of 1–3 × 10⁸ transformants/μg DNA. Transformation frequency was dependent on the electrical field strength and the pulse length. No significant reduction in transformation efficiency was observed when the transforming DNA contained sites sensitive to endonuclease $Atu CI$ of A. tumefaciens.

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Therefore, at least in silico, this strain appears to encode a functional RM system that might limit the transformation efficiency. Finally, as already mentioned, in Agrobacterium ID135 that also codes for a putative RM system, methylated DNA has been reported to be more efficiently transformed than unmethylated DNA (Mersereau et al., 1990). Consequently, methylome data of Ach5 would be very informative to uncover whether 5′-CCWGG-3′ is indeed methylated in the genome and, if so, removal of restriction recognition sites in a given plasmid might be useful to improve transformation efficiencies.
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Gene

Abstract

References (15)

Specificity of restriction endonucleases and methylases — a review (Edition 2)

Gene

Gene transfer in plants: production of transformed plants using Ti plasmid vectors

Methods Enzymol.

The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers

Gene

Agrobacterium DNA and PS8 bacteriophage DNA not detected in crown gall tumors

Properties of a deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event

Biochemistry

Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium melilotti

High efficiency transformation of E. coli by high voltage electroporation

Nucleic Acids Res.

Cited by (143)

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Agrobacterium strains and strain improvement: Present and outlook

Establishment of an efficient callus transient transformation system for Vitis vinifera cv. ‘Chardonnay’

The Arabidopsis thaliana onset of leaf death 12 mutation in the lectin receptor kinase P2K2 results in an autoimmune phenotype

Overexpression of the cotton trihelix transcription factor GhGT23 in Arabidopsis mediates salt and drought stress tolerance by binding to GT and MYB promoter elements in stress-related genes

Short communicationEfficient transformation of Agrobacterium tumefaciens by electroporation

Abstract

Gene

Methods Enzymol.

Gene

Agrobacterium DNA and PS8 bacteriophage DNA not detected in crown gall tumors

Properties of a deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event

Biochemistry

Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium melilotti

High efficiency transformation of E. coli by high voltage electroporation

Nucleic Acids Res.

Short communication
Efficient transformation of Agrobacterium tumefaciens by electroporation