Construction of a genetically engineered microorganism for CO2 fixation using a Rhodopseudomonas/Escherichia coli shuttle vector.
The CO2 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 PstI 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.
Du
Cuihong
C
School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian 116023, PR China. ducuihong@sohu.com
Zhou
Jiti
J
Wang
Jing
J
Yan
Bin
B
Lu
Hong
H
Hou
Hongman
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D002245:Carbon Dioxide / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005818:Genetic Engineering; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D012241:Rhodopseudomonas / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D002245:Carbon Dioxide; D012273:Ribulose-Bisphosphate Carboxylase