title: [Construction of the coexpression vector containing key element GLCYP450 involved in Ganoderma triterpene biosynthesis and its reductase gene GLNADPH].
abstract: Cytochrome P450 (CYP450) is a key element in the Ganoderma triterpenoid biosynthetic pathway. The catalytic reaction process for CYP450 requires NADPH / NADH for electron transfer. After searching the genome dataset of Ganoderma lucidum, the unique sequence encoding CYP450 and NADPH were discovered, separately. The open reading frames of GLCYP450 and GLNADPH were cloned separately using RT-PCR strategy from G lucidum. The appropriate restriction enzyme cutting sites were introduced at the 5' and 3' ends of gene sequence. The genes of GLCYP450 and GLNADPH were recombined into the yeast expression vector pESC-URA, leading to the formation of the yeast expression plasmid pESC-GLNADPH-GLCYP450. This study provides a foundation for researching Ganoderma triterpene biosynthesis using the approach of synthetic biology.
authors.affiliation: National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Guo
authors.forename: Chao
authors.initials: C
authors.lastname: Sun
authors.forename: Jing-Yuan
authors.initials: JY
authors.lastname: Song
authors.forename: Hong-Mei
authors.initials: HM
authors.lastname: Luo
authors.forename: Shi-Lin
authors.initials: SL
authors.lastname: Chen
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors; D009249:NADP / Q000235:genetics*; D016366:Open Reading Frames; D010957:Plasmids; D020940:Reishi / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D058615:Synthetic Biology; D014315:Triterpenes / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D014315:Triterpenes; D009249:NADP; D003577:Cytochrome P-450 Enzyme System