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Cloning of the Gene Encoding FNR Domain of FNR of Cyanobacterium Synechococcus sp. PCC 7002 and its Overexpression in Escherichia coli
Authors:LI Rong-Gui  ZHAO Jin-Dong  WU Guang-Yao  WU Xiang-Yu
Abstract:The petHL of Synechnococcus sp. PCC 7002 encoding FNR domain (FNRD) of FNR was amplified by PCR, and cloned into expressing vector pET-3a. Ovemxpression of petHL was achieved with E. coli BL21 (DE3). The recombinant FNRD was purified to homogeneity by DEAE-Sephadex A-50 and Sephadex G-100 chromatography. N-terminal amino acid sequencing showed that rFNRD was encoded by petHL and initial Met was not posttranslationally removed, rFNRD had the same absorption spectrum, optimal pH and optimal temperature as those of rFNR. rFNRD could catalyze photosynthetic electron transport from PT00 to NADP+ in vitro.
Keywords:Cyanobacteria   FNR domain   Overexpression   Electron transport  
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