Actinobacillus succinogenes shuttle vector and methods of use

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound

Reexamination Certificate

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C435S476000, C435S320100

Reexamination Certificate

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10911961

ABSTRACT:
AnActinobacillus succinogenesplasmid vector which provides a means to overexpress proteins inA. succinogenes. The plasmid can be transformed efficiently by electroporation, and replicates in a stable manner inA. succinogenes. The plasmid comprises at least one marker gene, operably linked to a first promoter functional inActinobacillus succinogenes, an origin of replication functional inActinobacillus succinogenes, a second promoter isolated fromActinobacillus succinogenes, and a cloning site downstream from the second promoter. Plasmids pLGZ901, pLGZ920, pLGZ921, and pLGZ922 are disclosed. The pckA gene polypeptide sequence and nucleic acid sequence ofActinobacillus succinogenes, including the promoter and ribosome binding site, is disclosed. Furthermore, a method for producing a recombinantActinobacillus succinogenesis described, including a method of transformation. Additionally, a recombinantActinobacillus succinogenesis disclosed and a method for producing succinate utilizing this recombinantActinobacillus succinogenesis described.

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