PCR Amplification of Tabersonine 16-Hydroxylase Gene From the Leaves of Catharanthus Roseus

Abstract

Hydroxylation of tabersonine by tabersonine 16-hydroxylase is a rate limiting step in the biosynthesis of terpenoid indole alkaloids (TIAs) in Catharanthus roseus. Total RNA was isolated from C. roseus leaves using three different methods of which isolation with TRIzol ® gave the best result. Phusion high-fidelity DNA polymerase amplified t16h gene from C. roseus cDNA while Taq polymerase failed due to high A-T content of t16h gene which needs proofreading DNA polymerase to increase the amplification efficacy.