[Molecular cloning of squalene synthase gene form Paris polyphylla and its expression in Escherichia coli].
OBJECTIVE
To clone the cDNA sequence of squalene synthase gene from Paris polyphylla, and characterize the biological features of the obtained SQS.


METHODS
Using homology cloning and RACE technique, a full-length cDNA sequence of PpSQS gene was isolated from P. polyphylla. The obtained sequence was analyzed by bioinformatics softwares. A plasmid [named pET-30b (+)-PpSQS] was constructed for prokaryotic expression the recombinant PpSQS.


RESULTS
The full-length cDNA of PpSQS gene is 1 498 bp, which contains a 1 212 bp ORF. Sequence analysis indicated that PpSQS encoded 403 amino acids residues with a calculated molecular weight (MW) of 46.36 kDa and an isoelectric point (pI) of 6.83. SDS-PAGE results showed that the recombinant PpSQS was expressed in Escherichia coli BL21 (DE3) by inducing with 1 mmol x L(-1) IPTG.


CONCLUSIONS
The full-length cDNA sequence of PpSQS gene was obtained from P. polyphylla, and its molecular features were consisted with classic SQS in plant. The recombinant PpSQS was successfully expressed in E. coli.
College of Life and Basic Sciences, Sichuan Agricultural University, Ya'an 625014, China. gaofeibiology@yahoo.cn
Fei
F
Gao
Xiao-Peng
XP
Luo
Liang
L
Tao
Cheng-Lei
CL
Li
Chun-Bang
CB
Ding
Hui
H
Chen
Qi
Q
Wu
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000235:genetics*; D019668:Liliaceae / Q000201:enzymology*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D013186:Farnesyl-Diphosphate Farnesyltransferase