Open AccessConstruction of a genetically engineered microorganism for CO 2 fixation using a Rhodopseudomonas / Escherichia coli shuttle vector
Author(s) -
Du Cuihong,
Zhou Jiti,
Wang Jing,
Yan Bin,
Lu Hong,
Hou Hongman
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00482-8
Subject(s) - multiple cloning site , shuttle vector , rhodopseudomonas palustris , biology , plasmid , ribosomal binding site , escherichia coli , microbiology and biotechnology , terminator (solar) , electroporation , expression vector , gene , molecular cloning , pbr322 , recombinant dna , gene expression , genetics , ribosome , vector (molecular biology) , bacteria , rna , ionosphere , physics , astronomy
The CO 2 fixation ability of Rhodopseudomonas palustris DH was enhanced by introducing the recombinant plasmid pMG‐CBBM containing the form II ribulose‐1,5‐bisphosphate carboxylase/oxygenase (RubisCO) gene ( cbbM ) isolated from Rps. palustris NO. 7. Sequencing of a 3.0‐kb Pst I fragment containing the cbbM gene revealed an open reading frame encoding 461 amino acids, homologous to known cbbM genes, with a ribosome binding site upstream of cbbM and a terminator downstream of cbbM , without promoter. pMG‐CBBM, a Rhodopseudomonas / Escherichia coli shuttle expression plasmid, was derived from the Rhodopseudomonas / E. coli shuttle cloning vector pMG105, by inserting the promoter of the pckA gene and the cbbM gene into its multiple cloning site. Plasmid pMG‐CBBM was transformed into Rps. palustris DH by electroporation, and was stably maintained when transformants were grown either photoheterotrophically or photolithoautotrophically in the absence of antibiotics. This is the first report of an expression plasmid containing a Rps. palustris ‐specific promoter that allows stable expression of a foreign gene in the absence of antibiotic selection.