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KMID : 0545120070170071106
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Journal of Microbiology and Biotechnology
2007 Volume.17 No. 7 p.1106 ~ p.1112
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Cloning and Characterization of Squalene Synthase (SQS) Gene from Ganoderma lucidum
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ZHAO MING-WEN
LIANG WAN-QI
ZHANG DA-BING
WANG NAN
WANG CHEN-GUANG
PAN YING-JIE
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Abstract
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This report provides the complete nucleotide sequences of the full-length cDNA encoding squalene synthase (SQS) and its genomic DNA sequence from a triterpene-producing fungus, Ganoderma lucidum. The cDNA of the squalene synthase (SQS) (GenBank Accession Number: DQ494674) was found to contain an open reading frame (ORF) of 1,404 bp encoding a 468-amino-acid polypeptide, whereas the SQS genomic DNA sequence (GenBank Accession Number: DQ494675) consisted of 1,984 bp and contained four exons and three introns. Only one gene copy was present in the G. lucidum genome. The deduced amino acid sequence of Ganoderma lucidum squalene synthase (Gl-SQS) exhibited a high homology with other fungal squalene synthase genes and contained six conserved domains. A phylogenetic analysis revealed that G. lucidum SQS belonged to the fungi SQS group, and was more closely related to the SQS of U. maydis than to those of other fungi. A gene expression analysis showed that the expression level was relatively low in mycelia incubated for 12 days, increased after 14 to 20 days of incubation, and reached a relatively high level in the mushroom primordia. Functional complementation of Gl-SQS in a SQS-deficient strain of Saccharomyces cerevisiae confirmed that the cloned cDNA encoded a squalene synthase.
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KEYWORD
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cDNA cloning, Ganoderma lucidum, squalene synthase, functional complement
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