A 1 250 bp cDNA fragment encoding β-mannanase, named MaMAN, was cloned from banana （Musa spp cv. Baxi） fruit using degenerate primers designed with reference to the conserved nucleic acid sequences of known β-mann...A 1 250 bp cDNA fragment encoding β-mannanase, named MaMAN, was cloned from banana （Musa spp cv. Baxi） fruit using degenerate primers designed with reference to the conserved nucleic acid sequences of known β-mannanase genes by RT-PCR. Sequence analysis showed that MaMAN cDNA encompassed a 1 085 bp open-reading frame （ORF）, encoding a predicted polypeptide of 395 amino acids. Alignment of the deduced amino acid sequence of MaMAN and other putative β-mannanases showed that MaMAN has an identity of 86, 70, 69, 54, and 57%, respectively, to β-mannanases from tomato, lettuce, arabidopsis, carrot and oryza sativa. The catalytic residues： Asn203, Glu204, Glu318 and the active site residues： Arg86, His277, Tyr279, and Trp360, which were strictly conserved in the glycoside hydrolase family 5 to which all β-mannanases belonged, were found in MaMAN. Semi-quantitative RT-PCR revealed that the level of MaMAN transcript in the pulp increased during banana fruit ripening, suggesting that MaMAN was likely to be involved highly in banana fruit softening.展开更多
文摘A 1 250 bp cDNA fragment encoding β-mannanase, named MaMAN, was cloned from banana （Musa spp cv. Baxi） fruit using degenerate primers designed with reference to the conserved nucleic acid sequences of known β-mannanase genes by RT-PCR. Sequence analysis showed that MaMAN cDNA encompassed a 1 085 bp open-reading frame （ORF）, encoding a predicted polypeptide of 395 amino acids. Alignment of the deduced amino acid sequence of MaMAN and other putative β-mannanases showed that MaMAN has an identity of 86, 70, 69, 54, and 57%, respectively, to β-mannanases from tomato, lettuce, arabidopsis, carrot and oryza sativa. The catalytic residues： Asn203, Glu204, Glu318 and the active site residues： Arg86, His277, Tyr279, and Trp360, which were strictly conserved in the glycoside hydrolase family 5 to which all β-mannanases belonged, were found in MaMAN. Semi-quantitative RT-PCR revealed that the level of MaMAN transcript in the pulp increased during banana fruit ripening, suggesting that MaMAN was likely to be involved highly in banana fruit softening.
