The gene encoding bacteriophage T7 RNA polymerase (T7gene1) was placed under the control of regulatory elements from the Escherichia coli lac operon to construct an inducible vaccinia virus expression system consisting entirely of prokaryotic transcriptional machinery. Regulated expression of T7 RNA polymerase was necessary to construct a stable recombinant vaccinia virus harboring a T7 promote...

Effective gene therapy requires efficient delivery of DNA to transfected cells followed by high levels of gene expression. Our laboratory has developed two new delivery formulations LPDI containing the DNA condensing agent protamine sulfate and reconstitued chylomicrons containing a hydrophobic lipidDNA complex. LPDI can produce higher levels of gene expression than the corresponding liposome/D...

A new Salmonella enterica serovar Typhimurium strain has been constructed to facilitate tightly regulated gene expression. Arabinose-inducible and glucose-repressible expression of a T7 RNA polymerase gene that has been integrated with an adjacent araC-P(BAD) control element into the bacterial chromosome allows dynamic control of T7 promoter-driven RNA transcription.

T7 gene 6 exonuclease has been shown to have an RNase H activity as well as a double-strand specific DNase activity by the following experiments: The RNase H activity coelutes with the DNase activity from DEAE-cellulose, phosphocellulose, hydroxyapatite, and Sephadex G-200 columns. Gene 6 exonuclease specified by a T7 strain with a temperature sensitive mutation in gene 6 has an extremely heat-...

The majority of the good DNA editing techniques have been developed in Escherichia coli; however, Bacillus subtilis is better host for a plethora of synthetic biology and biotechnology applications. Reliable and efficient systems for the transfer of synthetic DNA between E. coli and B. subtilis are therefore of the highest importance. Using synthetic biology approaches, such as streamlined lamb...

Caulobacter crescentus RNA polymerase holoenzyme and core enzyme have been separated by chromatography on denatured DNA-cellulose and purified by phosphocellulose chromatography. In order to assess the functional role of the putative C. crescentus CT subunit, the enzymes were compared to Escherichia coli holoenzyme and core RNA polymerases, and the transcription of various DNA templates by C. c...