Molecular cloning and expression analysis of a MGAPDH homologous gene from mango.
Luo Cong, He XinHua, Hu Ying, Tan Chao, Ou ShiJin
Author Affiliation: College of Agriculture, Guangxi University, Nanning 530004, China.
Journal of Fruit Science 28 : 1019-1024
Abstract : Glyceral-dehyde-3-phosphate dehyrogenase (GAPDH) is a key enzyme in sugar metabolism and energy metabolism. PCR primers were designed by the conserved sequences of GAPDH gene, and a full-length cDNA sequence of MGAPDH homologous gene was firstly cloned from Mango by employing RT-PCR and modified RACE technique. The full-length cDNA is 1 356 bp and contains an ORF with 1 203 bp, corresponding to a deduced protein of 401 amino acids. The results of bioinformatics analysis showed that the estimated molecular weight and isoelectric point of the putative protein were 42.8 ku and 8.9. The protein had two conserved domains, named as NADB_Rossmann superfamily and Gp_dh_C superfamily. Bioinformatics analysis also indicated that the protein of mango MGAPDH is a non-secreting protein without any signal peptide and transmembrane domain, MGAPDH gene was located in the chloroplast. The phylogenetic analysis based on the sequence of amino acids showed that MGAPDH protein may relative to photosynthesis. Semiquantitative analysis showed that the MGAPDH homologous gene expressed in all tissues of mango, no significant differences in expression levels were founded among the tested tissues; the result indicted that MGAPDH homologous gene may be used as an internal standard for differential gene expression analysis in mango.