Cloning of ATP binding protein gene in sugarcane and its expression characteristics under drought stress

Objective The present experiment was conducted to clone ATP binding protein gene in sugarcane,analyze its bioinformatics and expression under drought stress,and provide reference for function research in ATP binding protein gene.MethodSequence of ATP binding protein gene was obtained from differential expression transcriptome induced by ethylene and sugarcane cDNA library under water stress.Its primers were amplified by RT-PCR,sequence analysis and protein structure and function prediction were carried out by bioinformatics software.Real-time fluorescence quantitative analysis was used to detect expression of sugarcane ATP binding protein gene under drought stress.ResultSugarcane ATP binding protein gene was cloned.It included 2145 bp and encoded 714 amino acids.Its relative protein molecular weight was 79.430 kD,isoelectric point(pI) 9.33 and instability index 44.38.The homology in nucleotide sequence and deduced amino acid sequence between sugarcane ATP binding protein gene and maize ATP binding protein gene was the highest (93 %).Sugarcane ATP binding protein gene mainly consisted of random coil (42.02%)、alpha helix (25.63 %) and extended strand (22.27%).Sugarcane ATP binding protein gene consisted of protein kinase catalytic domain,ATP binding site,nucleotide binding site (NBS),glycosylation site,kinase phosphorylation binding site and nuclear localization signal.The results of subcellular localization and protein function classification prediction showed that sugarcane ATP binding protein might be mainly located in nucleus,chloroplast,mitochondria and peroxidase,and played a role of intermediary metabolism.It might involved in biological reactions such as transcription regulation,immune response,stress response and signal transduction.The results of real-time qPCR analysis showed that,7 d after drought stress,the expression of sugarcane ATP binding protein gene down regulated to 30% ofthat in control,and after returning to normal water supply,the expression was on the rise.conclusionSugarcane ATP binding protein gene is induced by drought stress and may be involved in metabolic process of sugarcane responsive to drought adversity stress.