Molecular cloning and in silico analysis of a GTP cyclohydrolase I gene from grape ‎

An entire open reading frame (ORF) encoding for a polypeptide of GTP cyclohydrolase I (GTPCH I) was isolated and cloned from Askari cultivar of grape (Vitis vinifera L.) berries. The ۱,۳۳۸-nucleotide ORF yields a ۴۴۵-residue amino acid sequence with a calculated molecular mass of ۴۸.۶۵ kDa and a predicted isoelectric point of ۶.۴۳. The Vvgtpch I genomic sequence with a length of ۴,۹۶۴ bp contains two exons (۱۶۹ and ۱,۱۶۹ bp) and an intron (۲,۶۷۶ bp). The gtpch I sequence of grape displayed a strong similarity with gtpch I sequence found in other plants, including peach (۷۲%), cocoa (۷۲%), strawberry (۷۰%), and poplar (۶۹%). Analysis of mRNA secondary structure revealed that the start codon of Vvgtpch I is completely exposed, suggesting a robust binding of the ribosome and efficient translation. Similar to gtpchs I from  diverse sources, molecular modeling uncovered that the monomer of VvGTPCH I adopts an αβ structure, which includes ۱۰ α-helices and ۸ β-sheets. Moreover, in silico analysis of the Vvgtpch I gene promoter identified potential cis-acting elements  responsive to environmental signals. This suggests that the Vvgtpch I gene has the capacity to be responsive to various environmental cues, such as heat, heavy metals, light, and plant hormones.

Keywords:

Folate , in silico analysis , Regulatory elements , Cloning , Promoter region