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Isolation and characterization of a gene encoding 3-hydroxy-3-methylglutary coenzyme A reductase from an endophytic taxol-producing fungus BT2

Wang, Yechun, Tang, Kexuan
Annals of microbiology 2012 v.62 no.2 pp. 587-595
Southern blotting, active sites, biochemical pathways, bioinformatics, complementary DNA, endophytes, exons, fungi, genes, introns, methyl jasmonate, open reading frames, paclitaxel, polypeptides, reverse transcriptase polymerase chain reaction
A full-length cDNA encoding 3-hydroxy-3-methylglutaryl-CoA reductase (designated as Bt2HMGR) and its genomic sequence were cloned and characterized from an endophytic taxol-producing fungus BT2 for the first time. The full-length cDNA of Bt2HMGR was 4,078 bp with an open reading frame of 3,522 bp encoding an 1,173-amino-acid polypeptide. The comparison analysis between cDNA and genomic DNA sequence revealed that Bt2HMGR gene had four exons and three introns. Bioinformatic analyses revealed that the deduced Bt2HMGR contained six transmembrane domains and a catalytic domain, and showed high similarity to other fungal HMGRs. Southern blot analysis indicated that Bt2HMGR was a single copy gene. RT-PCR analysis indicated that the expression of Bt2HMGR could be induced by methyl jasmonate acid. The cloning and characterization of Bt2HMGR will enable us to further understand the role of Bt2HMGR in the taxol biosynthetic pathway in an endophytic taxol-producing fungus.