Author/Authors :
Basherudin, Norlia Forest Research Institute Malaysia - Genetic Unit, Biotechnology Division, Malaysia , Muhammad, Norwati Forest Research Institute Malaysia (FRIM) - Genetic Unit, Biotechnology Division, Malaysia , Adnan, Norwati Forest Research Institute Malaysia (FRIM) - Genetic Unit, Biotechnology Division, Malaysia , Haron, Mohd Rosli Forest Research Institute Malaysia (FRIM) - Genetic Unit, Biotechnology Division, Malaysia , Saleh, Norihau Mohd Universiti Putra Malaysia - Faculty of Biotechnology and Biomolecular Science - Cell and Molecular Biology Department, Malaysia
Abstract :
Expression of an isolated gene in a system that directly translates it into a protein is an important step to study the protein encoded by the gene. The isolated gene can be expressed in vivo by a heterologous system. In this study, a bacteria system was used to translate the Tectono grandis Late Elongated Hypocotyl (Tg-LHY) gene, which was isolated from flowering tissues of teak (Tectona grandis). The gene was cloned into the pET 14b vector (Novagen) and transformed into BL 21 (DE3) /pLysS and Rosetta 2 expression host cells (Novagen). Rosetta 2 host cell has been found to be a good candidate to express the Tg-LHY protein from plant origin, as it recognizes the codon that was found in plant but rarely used in bacteria. The expressed protein was about an expected size , which was 90 kD. Western blot analysis using antibody against His-tag, which was fused to the Tg-LHY protein, proved that the expressed protein was Tg-LHY protein.
