원문정보
초록
영어
Cyanobacteria are photoautotrophic organisms that can assimilate CO2 into organic materials
using photon energy. They also evolve O2 gas from water splitting during photosynthesis. Due to
these prominent features, there are potentials to exploit them for clean, renewable fuel production,
especially biohydrogen. To improve cyanobacterial strain for efficient hydrogen production,
genetic manipulation technique is needed. Thus, in the present work, we constructed a shuttle
expression vector for Synechocystis PCC 6803 as a first footstep. It consists of three parts which
came from three different plasmids. The replication origin (oriV), origin of transfer (oriT), and
associated genes (repA,B,C ) were cloned as a 5.6kbp fragment from RSF-1010, a broad host range plasmid, allowing to replicate autonomously in some cyanobacterial strains including
Synechocystis PCC 6803. The commercial vectors, pACYCDuet-1 (Novagen) and pTrcHisC
(Invitrogen), provided chloramphenicol acetyl transferase (cat) gene, Trc promoter, multiple
cloning sites (MCS), and other accessory sequences. The latter two parts were PCR-cloned. We
introduced green fluorescent protein (GFP) gene into the MCS region of the 7.2kbp-long
complete vector which was named pRCT. We will present functional expression of reporter GFP in E. coli and Synechocystis PCC 6803 strains to confirm successful construction.