Molecular cloning and in silico analysis of DREB-like gene in watermelon

Abstract

The objective of this study was to identify and characterize sequences from watermelon related to drought tolerant genes. In order to achieve this, publicly available Cucumis melo REB gene (Cme-DREB1) sequence was retrieved from NCBI database and gene-specific primers were synthesized. A fragment of 274 bp corresponding to DREB-like gene was amplified from genomic DNA of watermelon. PCR product was cloned in E. coli vector (pDrive) and validated through sequencing. The BLAST search showed 99 and 97% homology to CmeDREB1 gene and C. sativus DREB3-like (CsDREB3-like) gene, respectively. Furthermore, it showed 87 to 93% homology with rest of DREB genes available in the public database. Full length watermelon DREB-like gene (C. lanatus DREB-like, ClaDREB-like) was identified from cucurbit genomic database and investigated through in silico analysis. The intronless structural organization of ClaDREB-like gene was found with a conserved AP2/ERF domain.