Molecular Cloning Of Three Oxidase Genes From Mulberry(Morus L.),Sequence Analysis And Expression

Plants during its own aerobic abiotic stress, the body will produce large amounts of reactive oxygen, such substances in plants if not promptly removed, will have serious toxic effects on plant growth and development. In order to maintain normal growth, plant clear reactive oxygen species and antioxidant enzyme system.China is the origincenter of mulberry, which use a lot of resources have value fruit, medicinal value, economic value and so on. Thus, to carry out the cloning of mulberry oxidase and peroxidase geneand functional verification and analysis, not only make us recognize the expression of mulberry oxidase and peroxidase gene under different periods and conditions from the molecular level, The more to understanding of its role and function in mulberry; and lay the foundation for future resistance mulberry obtain different environments and access to high-yield high-quality mulberry resources is a great significance.1、Mulberry(Morus multicaulis) kaurene oxidase gene cloning research and their expression analysis and sequence analysis.By mulberry cDNA library and software applications and a variety of experimental techniques, get the first mulberry kaurene oxidase gene fragment of 1838 bp and named MmKO, contains an open reading frame length of 1551 bp, encoding 516 amino acids protein, predicted mulberry kaurene oxidase protein molecular mass 58.563 kD, isoelectric point of 7.917, belongingto p450 superfamily. And with other species encoded amino acid oxidase kaurene homology comparison and gene sequences from other species kaurene oxidase conducted phylogenetic analysis. PCR results showed that, compared with the normal growth environment, MmKO gene in drought, salt, ABA, SA four abiotic environmental stress conditions, regulating the expression levels of all changes at different levels.2、Molecular cloning,Sequence analysis and induced expression studies of peroxidase 12 from mulberry(Morus multicaulis).By mulberry cDNA library and software applications and a variety of experimental techniques, get some coding a sequencemulberry peroxidase 12(MmPOD12) gene, a cDNA length 1482 bp, one complete open reading frame containing the complete open reading frame the sequence length is 789 bp, encoding the amino acid residue corresponding to the number of 350. Predict protein molecular mass of 37.004 kD, an isoelectric point of 7.044,belonging to peroxidase gene family. Homology in amino acid alignments, MmPOD12 gene has a certain conservative among the species, there are differences, and Morus notabili maintain relatively close genetic relationship. Fluorescence quantitative PCR: MmPOD12 gene expression levels under drought, salt, ABA, SA stress conditions will produce some changes in the regulation.3、Molecular cloning,sequence analysis and induced expression studies of primary amine oxidase 2 from mulberry(Morus multicaulis).By mulberry cDNA library and software applications and a variety of experimental techniques have been constructed, mulberry primary amine oxidase obtained 2 contains a complete open reading frame of cDNA length 2597 bp, open reading frame sequence containing more complete and the length of 2364 bp, the number of encoded protein is 787 amino acids. The predict two primary amine oxidase protein molecular mass is 87.53 kD, an isoelectric point of 6.545, belonging to copper amine oxidase gene family. Utilizing homology software PAO2 mulberry and other species a gene encoding the amino acid build alignments and phylogenetic tree; the results obtained by quantitative PCR revealed that the expression level MmPAO2 gene mRNA compared to normal growth environment, which in drought, salt, ABA, SA will have four non-regulation of the expression of some movements in the biological environment under stress conditions.