Molecular cloning and functional expression analysis of a new gene encoding geranylgeranyl diphosphate synthase from hazel (Corylus avellana L. Gasaway).
Geranylgeranyl diphosphate synthase (GGPPS) [EC 2.5.1.29] catalyzes the biosynthesis of geranylgeranyl diphosphate (GGPP), which is a key precursor for diterpenes such as taxol. Herein, a full-length cDNA encoding GGPPS (designated as CgGGPPS) was cloned and characterized from hazel (Corylus avellana L. Gasaway), a taxol-producing angiosperms. The full-length cDNA of CgGGPPS was 1515 bp with a 1122 bp open reading frame (ORF) encoding a 373 amino acid polypeptide. The CgGGPPS genomic DNA sequence was also obtained, revealing CgGGPPS gene was not interrupted by an intron. Southern blot analysis indicated that CgGGPPS belonged to a small gene family. Tissue expression pattern analysis indicated that CgGGPPS expressed the highest in leaves. RT-PCR analysis indicated that CgGGPPS expression could be induced by exogenous methyl jasmonate acid. Furthermore, carotenoid accumulation was observed in Escherichia coli carrying pACCAR25ΔcrtE plasmid carrying CgGGPPS. The result revealed that cDNA encoded a functional GGPP synthase.
Wang
Yechun
Y
Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, 200240, Shanghai, China.
Miao
Zhiqi
Z
Tang
Kexuan
K
D000085:Acetates / Q000494:pharmacology; D001483:Base Sequence; D015139:Blotting, Southern; D002338:Carotenoids / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology; D031211:Corylus / Q000201:enzymology* / Q000235:genetics*; D003517:Cyclopentanes / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D051231:Farnesyltranstransferase / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005816:Genetic Complementation Test; D018745:Genome, Plant / Q000235:genetics; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000494:pharmacology; D015183:Restriction Mapping
D000085:Acetates; D003517:Cyclopentanes; D018076:DNA, Complementary; D054883:Oxylipins; D002338:Carotenoids; C072239:methyl jasmonate; D051231:Farnesyltranstransferase