Journal of Pure and Applied MicrobiologyVol. 8 No. May 2014 Special Edition

Molecular Cloning and Expression of SS Gene from Panax japonicus

Lai Zhang1,2* and Min Sun2

1School of Life Science, Southwest University , Key Laboratory of Eco-environments in Three Gorges Reservoir Region(MOE) Chongqing 400715 China. 2College of Agriculture?Anshun University/Key lab of Function Material and Resource Chemistry (GZSJYT), An shun 561000 Gui zhou, China.

Received on 12 April 2014 and accepted on 09 May 2014

 

ABSTRACT

The squalene synthase (SS) gene from Panax japonicus is an important regulatory enzyme in the biosynthetic pathway of triterpenoid saponins (TS). Regulating the SS activity directly affects the synthesis of squalene (SQ), as well as the synthesis of TS, for which SQ acts as a precursor. By using the homologous cloning program RACE, the SS in the synthetic TS pathway of P. japonicus was successfully cloned , it was named for PjSS. Its fragment was 1353 bp, encoding 415 amino acids. The PjSS protein was an unstable and hydrophobic protein, with the molecular mass of 109559.7 kD. The secondary structure of SS was composed of a-helix, random coil, extended strand, and b-angle of 67.47%, 22.41%, 7.23%, and 2.89%, respectively. The PjSS protein exhibited folding properties to form the typical three-dimensional structure,which has eight functional motifs.RT-PCR indicated that the expressing quantity of PjSS is max in the hypogaeic tuber, the root is second, and the leaf is min.

Keywords : Squalene synthase gene, Panax japonicus, Cloning, Expression.