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Ralstonia eutropha H16 (also known as Cupriavidus necator H16) is a Gram‐negative lithoautotrophic β‐proteobacterium with increasing biotechnological applications, including carbon capture and utilization, biopolymer synthesis and biofuel production. Engineering of this organism is supported by the availability of its genome sequence and suitable plasmid systems. However, the lack of a simple and robust transformation method remains a challenge as it limits both the pace and ease of engineering this organism. To overcome this limitation, a systematic study was performed to evaluate the effects of different parameters on the transformation efficiency of R. eutropha H16. The optimized electroporation protocol uses R. eutropha H16 cells grown to OD600 0.6. These cells were made competent by a 15‐min incubation in 50 mM CaCl2, followed by two cell washes and final resuspension in 0.2 M sucrose prior to electroporation using 2.3 kV. This protocol achieved a transformation efficiency of (3.86 ± 0.29) x 105 cfu/μg DNA, a 103‐fold improvement compared to a previously published value for the same plasmid. This transformation method is a valuable tool for R. eutropha H16 research and will further enable the development of other advanced molecular biology methods for this industrially relevant microorganism.NEXT ARTICLE
Antibodies Antisense Assays Biochips Bioinformatics Biological Therapy Biomarkers Biomaterials Bioscience Cell Culture Cloning Cytokine Diagnostics Dna Extraction Dna Sequencing Dna Transform...
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...