Host-vector system for cloning and expressing genes
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United States of America Patent
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Jan 7, 2014
Grant Date -
Oct 7, 2010
app pub date -
Apr 6, 2010
filing date -
Apr 6, 2009
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Abstract
A system for ligase-free cloning and/or expressing a target gene is described herein. A preferred version of the invention includes an E. coli host. The host preferably includes a T7 RNA polymerase gene comprising a T7gpl coding sequence, a lacUV5 promoter, and a lac operator. The host preferably further includes a lacI gene comprising a lacI coding sequence with an ATG start codon, a promoter derived from the lacql allele, and a translational enhancer derived from a 5′ RNA leader sequence of T7 gene 10. The invention further includes a low-copy plasmid vector comprising a T7 promoter a lac operator operationally linked to the T7 promoter. The system is configured to inhibit target gene expression when uninduced and to permit gene expression upon induction by auto-induction.
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