In Silico Cloning and Sequence Analysis of F3H Gene in Phaseolus coccineus
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Abstract:
Flavanone 3-hydroxylase is a key enzyme in the biosynthetic pathway of plant flavonoids. The putative cDNA sequence of Phaseolus coccineus F3H gene was in silico cloned by using Glycine max F3H gene cDNA sequence (AAU06216.1) as a probe. According to the putative cDNA, the primers were designed for RT-PCR. The coding sequence of F3H gene in Phaseolus coccineus cDNA was cloned using RT-PCR. Then the secondary structure and advanced structure of F3H in Phaseolus coccineus were analyzed by bioinformatics methods. The results showed that the cDNA contains an open reading frame of 1128 bp, which encoded the protein containing 375 amino acids. The protein coded by Phaseolus coccineus F3H gene cDNA showed 92% similarity to Glycine max. Its secondary structure contained 40.8% α-helix, 14.93% extended strand, 39.73% random coil and 4.53% β-turn. The homology analysis of three-dimensional structure showed that the three-dimensional structure of the protein was a compact globular structure.
