Glyceraldehyde-3-phosphate Dehydrogenase Gene from Halophyte Aeluropus lagopoides: Identification and Characterization

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which plays an important role in central carbon metabolism, was studied in Aeluropus lagopoides, which is an important perennial grass found in drought-stricken areas and deserts from Africa to South Asia. Aeluropus lagopoides is an ideal candidate plant system to identify novel or enhanced stress tolerance mechanisms. The GAPDH has stable expression under different conditions and it is a typical housekeeping gene in prokaryotes and eukaryotes. So far, there are no reports about identification of housekeeping genes for gene expression analysis in Aeluropus lagopoides. Thus in the present study, a GAPDH cDNA was cloned and sequenced from this plant. Our results revealed that the open reading frame (ORF) (1014 bp) encoded a polypeptide of 337 amino-acid residues. The deduced amino acid sequence indicated a protein with a molecular weight of 364.95 kDa and estimated isoelectric point value of 7.06. Multiple alignments of putative amino acid sequences indicated high similarity between Aeluropus lagopoides GAPDH and analogous protein derived from other plants, particularly monocots. Nucleotide blast and phylogenetic analysis revealed that this plant had maximum identity with Cleistogenes songorica. Results of this experiment can help us do transcriptional analysis by qRT-PCR to identify stress-tolerance mechanisms.

Keywords:

• cDNA
• gene cloning
• housekeeping gene

Keywords:

• (EF1alpha) elongation factor 1-alpha
• (IP) iesoelectric point
• (NAD⁺) nicotinamide adenine dinucleotide
• (qRT-PCR) quantitative real-time reverse transcription polymerase chain reaction.