Isolation, characterization and expression analysis of a leaf-specific phosphoenolpyruvate carboxylase gene in Oryza sativa.
Suppression subtractive hybridization was carried out to enrich gene fragments over-expressed in rice leaves by subtraction to rice roots, from which two identical cDNA fragments were identified to encode putative phosphoenolpyruvate carboxylase. Then the corresponding full-length cDNA (Osppc) is isolated by RT-PCR and sequenced, which indicates an open reading frame of 2895bp is contained. Its deduced protein is encoded in 10 exons and shows high similarity to many other plant PEPCs. Comparing with maize and bacterial PEPCs, it is revealed that OSPPC shares many conserved domains and active sites that responsible for the structure, activity and regulation of this enzyme. Phylogenetic analysis demonstrates that OSPPC is grouped with C3 form PEPCs of wheat, maize and sorghum, which is consistent with the classification of rice. And a putative promoter element is predicted with DOF binding box, CAAT box and TATA box in the 5'-flanking sequence of Osppc gene. Moreover, Quantitative RT-PCR analyses are performed in hybrid rice and its parents, which show that Osppc is specifically expressed in leaf including leaf vein and sheath.
Lin
Chang-Fa
CF
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, PR China.
Wei
Chun
C
Jiang
Li-Zhi
LZ
Li
Ke-Gui
KG
Qian
Xiao-Yin
XY
Attia
Kotb
K
Yang
Jin-Shui
JS
D000595:Amino Acid Sequence; D001483:Base Sequence; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D010730:Phosphoenolpyruvate Carboxylase / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D010730:Phosphoenolpyruvate Carboxylase