Abstract
Maintenance of recombinant plasmid vectors in host bacteria relies on the presence of selection antibiotics in the growth media to suppress plasmid -free segregants. However, presence of antibiotic resistance genes and antibiotics themselves is not acceptable in several applications of biotechnology. Previously, we have shown that FabV-Triclosan selection system can be used to select high and medium copy number plasmid vectors in E. coli. Here, we have extended our previous work and demonstrated that expression vectors containing FabV can be used efficiently to express heterologous recombinant proteins in similar or better amounts in E. coli host when compared with expression vectors containing β-lactamase. Use of small amount of non-antibiotic Triclosan as selection agent in growth medium, enhanced plasmid stability, applicability in various culture media, and compatibility with other selection systems for multiple plasmid maintenance are noteworthy features of FabV-Triclosan selection system.
Highlights
Plasmid vectors frequently employ antibiotic resistance genes as selection markers; growth media are regularly supplemented with antibiotics. [1]
We have extended our previous work [18] and demonstrated that fabV-Triclosan selectable medium copy number plasmid vectors can be used efficiently to express heterologous recombinant proteins in similar or better amounts in E. coli when compared with traditional bla (β-lactamase)-Ampicillin selectable vectors
This report is an extension of our previous work where we have shown that overexpression of V. cholerae fabV gene can be used to select pBR322 -based medium copy plasmid vectors in culture medium supplemented with Triclosan [18]
Summary
Plasmid vectors frequently employ antibiotic resistance genes as selection markers; growth media are regularly supplemented with antibiotics. [1]. Plasmid vectors frequently employ antibiotic resistance genes as selection markers; growth media are regularly supplemented with antibiotics. For large-scale fermentation, plasmid vector selection using antibiotics can prove expensive [3]. Disadvantages associated with antibiotic -based selection have encouraged development of antibiotic marker -free selection approaches. Several such systems have been reported [4, 5, 6, 7, 8, 9, 10, 11] but not adopted commonly by research laboratories for numerous reasons that, among other, include use of specialized strains and/or reagents.
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