Exon prediction, protein prediction and functional analysis

The identified partial sequence of LprPHYA3 was subjected to exon prediction using Eukaryotic GeneMark.hmm version 3.54 [36]. The nucleotide sequence of exon-1 obtained in such a way was translated to protein sequence using standard codon table. This amino acid sequence obtained was further compared with GmPHYA3 sequence. The protein sequence of exon 1 from Lablab purpureus GNIB-21 was used to perform BLASTp in NCBI GenBank database and the sequences showing homology were further used to create multiple alignment using the CLUSTAL W algorithm [29, 31]. Phylogenetic analysis with amino acid sequence of PHYA in 21 different plant species was also performed using JTT model in MEGA X [30, 37]. The online web-based functional analysis tool SMART-EMBL (Simple Modular Architecture Research Tool) (http://smart.embl-heidelberg.de/) Version 9 was used for predicting domains in both GmPHYA3 and LprPHYA3 protein sequences [38].