Molecular cloning and identification of tissue-specific expression of ent-kaurene oxidase gene in Momordica charantia

Abstract

Momordica charantia (bitter melon) is widely consumed as a vegetable and as a folk medicine. Ent-kaurene oxidase is a key enzyme of gibberellin (GA) synthesis by controlling the early GA biosynthesis. In this study, ent-kaurene oxidase cDNA sequence was successfully amplified from the total cDNA of pistillate flower buds, and a complete open reading frame (ORF) with 1563 bp, encoding 520 amino acids, containing 151 bp of 5’-untranslated region sequences and 217 bp of the 3’-untranslated region sequences was found. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that the expression level of M. charantia ent-kaurene oxidase (McEKO) gene in pistillate flowers was significantly higher than in staminate flowers and neutral flowers, and the expression level was especially abundant in root and stem apexes. The gene was express in Escherichia coli cells with a molecular weight of 58.8 kD observed.

Key words: Gibberellin, bioinformatics analysis, procaryotic expression, quantitative real-time polymerase chain reaction qRT-(PCR).