Abstract: Cyclotides are highly tolerant of sequence variability, aside from the conserved residues forming the cystine knot. Cyclotides have attractive natural movement capabilities including cytotoxicity, harmfulness to HIV, antimicrobials, insecticides, uterine contraction and inhibition of proteases, making them very important in pharmacy and horticulture. The medicinal cyclotides may be able to reimburse the safety of antimicrobials, which is a noticeable problem worldwide today. The present study was focused on the isolation of genes that encoding cyclotide, from plant species. DNA was isolated by CTAB method. The gene was amplified by PCR and after agarose gel electrophoresis confirmation that gene was sequenced followed by bioinformatics analysis. Petunia showed 90% to 97% identity with cds of reference sequence and 27 amino acids were identical in multiple alignment of protein sequence. In addition to providing information for the future investigation of Solanaceae species for cyclotides, this work will aid in understanding the plant families that produce cyclotides.