[Cloning and sequence analysis of squalene synthase gene and cDNA in Glycyrrhiza uralensis].
OBJECTIVE
To clone and sequence the open reading frame and genomic sequence of squalene synthase (SQS) from Glycyrrhiza uralensis.


METHODS
The primers were designed according to cDNA sequence of SQS from G. glabra reported by Hiroaki HAYASHI, SQS cDNA was cloned with total RNA extracted from roots of G. uralensis. Specific fragments were amplified by RT-PCR and then were cloned and sequenced. SQS DNA was cloned with total DNA extracted from roots of G. uralensis. Specific fragments were amplified by PCR and then were cloned and sequenced.


RESULTS
GuSQS1 (GenBank accession number: GQ266154) was 1 242 bp in length encoding proteins with 412 amino acid. NCBI Blast x search results showed GuSQS1 had the highest amino acid similarity to the corresponding proteins from G. uralensis. The identities of GuSQS1 with the two proteins were 98. 55% and 88. 62%. SQS (GenBank accession number: GQ180932) gene with 4 484 bp containing 13 exons and 12 introns was then amplified by PCR with genomic DNA extracted from roots of G. uralensis.


CONCLUSIONS
These findings of cloning and sequencing the open reading frame and genomic sequence of squalene synthase (SQS) from G. uralensis brought some new clues for the further exploration of SmSQS function in sterol and terpenes biosynthesis.
School of Chinese Materia Medica, Beijing University of Traditional Chinese Medicine, Beijing 100102, China. rqx1982@126.com
Qixian
Q
Rong
Chunsheng
C
Liu
Luqi
L
Huang
Ning
N
Zhang
Bo
B
Nan
Wei
W
Guo
D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000737:chemistry; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000737:chemistry*; D029882:Glycyrrhiza uralensis / Q000737:chemistry* / Q000201:enzymology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D018517:Plant Roots / Q000737:chemistry / Q000201:enzymology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods
D018076:DNA, Complementary; D013186:Farnesyl-Diphosphate Farnesyltransferase