Molecular cloning and characterization of a novel stem-specific gene from Camptotheca acuminata.
In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissuespecific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.
Pi
Yan
Y
Institute of Genetics, School of Life Sciences, Morgan-Tan International Center for Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Fudan University, Shanghai 200433, People's Republic of China.
Liao
Zhihua
Z
Chai
Yourong
Y
Zeng
Hainian
H
Wang
Peng
P
Gong
Yifu
Y
Pang
Yongzhen
Y
Sun
Xiaofen
X
Tang
Kexuan
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D029769:Camptotheca / Q000033:anatomy & histology / Q000201:enzymology / Q000235:genetics; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018514:Plant Structures / Q000201:enzymology* / Q000235:genetics; D010946:Plants, Medicinal / Q000033:anatomy & histology / Q000201:enzymology / Q000235:genetics; D011487:Protein Conformation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D010940:Plant Proteins; D003577:Cytochrome P-450 Enzyme System