Mutants in orthophosphate-regulated cyclic phosphodiesterase showing rhythmic condiation in Neurospora

Mutants in orthophosphate-regulated cyclic phosphodiesterase showing rhythmic conidiation in Neurospor

A simple method to isolate mutants in repressible cyclic phosphodiesterase in Neurospora crassa

A simple method to isolate mutants in repressible cyclic phosphodiesterase in Neurospora crassa

An efficient isolation method for meiotic mutants causing meiotic nondisjunction or elevated recombination frequency in Neurospora crassa.

An efficient isolation method for meiotic mutants causing meiotic nondisjunction or elevated recombination frequency in Neurospora crassa

An efficient isolation method for polyadenylated messenger ribonucleic acid from Neurospora mycelia

An efficient isolation method for polyadenylated messenger ribonucleic acid from N. crassa

Assay method and localization of GTP binding proteins in N. crassa.

Assay method and localization of GTP binding proteins in N. crassa

A method to analyze the mode of action of hormones using conidiation rhythm of Neurospora.

A method to analyze the mode of action of hormones using conidiation rhythm of Neurospora

Stable Maintenance of Multiple Plasmids in E. coli Using a Single Selective Marker

Plasmid-based genetic systems in Escherichia coli are a staple of synthetic biology. However, the use of plasmids imposes limitations on the size of synthetic gene circuits and the ease with which they can be placed into bacterial hosts. For instance, unique selective markers must be used for each plasmid to ensure their maintenance in the host. These selective markers are most often genes encoding resistance to antibiotics such as ampicillin or kanamycin. However, the simultaneous use of multiple antibiotics to retain different plasmids can place undue stress on the host and increase the cost of growth media. To address this problem, we have developed a method for stably transforming three different plasmids in E. coli using a single antibiotic selective marker. To do this, we first examined two different systems with which two plasmids may be maintained. These systems make use of either T7 RNA polymerase-specific regulation of the resistance gene or split antibiotic resistance enzymes encoded on separate plasmids. Finally, we combined the two methods to create a system with which three plasmids can be transformed and stably maintained using a single selective marker. This work shows that large-scale plasmid-based synthetic gene circuits need not be limited by the use of multiple antibiotic resistance genes