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Design of a microaerobically inducible replicon for high-yield plasmid DNA production.

Biotechnol. Bioeng.. 2019-06; 
JaénKarim E,VelázquezDaniela,SigalaJuan-Carlos,LaraAlva
Products/Services Used Details Operation
Plasmid DNA Preparation … USA Inc. (Piscataway, NJ, USA). Constructions of the plasmids were verified by restriction analysis and Sanger sequencing of the inserts by GenScript USA Inc. and Unit of DNA Synthesis and Sequencing (Universidad Nacional Autónoma de México) … Get A Quote

摘要

A pUC-derived replicon inducible by oxygen limitation was designed and tested in fed-batch cultures of Escherichia coli. It included the addition of a second inducible copy of rnaII, the positive replication control element. The rnaII gene was expressed from P and cloned into pUC18 to test the hypothesis that the ratio of the positive control molecule RNAII to the negative control element, RNAI, was the determinant of plasmid copy number per chromosome (PCN). The construct was evaluated in several E. coli strains. Evaluations of the RNAII/RNAI ratio, PCN and plasmid yield normalized to biomass (Y ) were performed and the initial hypothesis was probed. Furthermore, in high cell-density cultures in shak... More

关键词

microaerobic induction,oxygen limitation,pUC replicon,plasmid DNA,r