Cloning and heterologous expression of a gene encoding lycopene-epsilon-cyclase, a precursor of lutein in tea (Camellia sinensis var assamica)

Abstract

This report describes the cloning and expression of a gene lycopene epsilon cyclase, (LCYE) from Camellia sinensis var assamica which is a precursor of the carotenoid lutein in tea. The 1982 bp cDNA sequence with 1599 bp open reading frame of LCYE was identified from an SSH library constructed for quality trait in tea. 5’ and 3’ RACE (rapid-amplification of cDNA ends) was done to clone the full length cDNA of LCYE. Homology studies showed that the deduced amino acid sequence of LCYE gene had the highest sequence identity of up to 84% with Vitis vinefera. The cloned gene was successfully expressed in a PET based Escherichia coli expression system. The size of the expressed protein was 59615 Daltons. A suppression subtractive library was constructed using a quality clone H3111 (tester) and a garden series clone T3E3 (driver).

Key words: Carotenoid, RACE, heterologous expression, lutein, tea.