N4 virion single-stranded DNA dependent RNA polymerase

Details N4 virion single-stranded DNA dependent RNA polymerase N4 virion single-stranded DNA dependent RNA polymerase

2002-05-22

2008-11-18

Hutson, Richard G (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

C435S183000, C435S252300, C435S320100, C435S006120, C536S023200

Reexamination Certificate

active

07452705

ABSTRACT:
A histidine-tagged, deletion mutant of bacteriophage N4-coded, virion RNA polymerase (mini-vRNAP) which is active has been developed. The his-tagged mini-vRNAP has been cloned under the control of the pBAD promoter, is stable and is purified in a single step yielding large amounts (10 mg/liter ofE. coliexpressing cells). This RNA polymerase uses single-stranded DNA containing 17 bases (the promoter) upstream of the transcribed regions as a template. In the presence ofE. coliSSB protein, it transcribes this template efficiently, providing a unique system to synthesize RNAs of the desired sequence using single-stranded DNA templates. The enzyme incorporates derivatized nucleoside triphosphates with high efficiency. A mutant of mini-vRNAP has been generated that incorporates deoxynucleoside triphosphates. In addition, the inventors have developed an in vivo system to express RNAs and proteins under mini vRNA polymerase promoter control.

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Database Accession No. ABG11811.

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