甘蔗s腺苷蛋氨酸脱羧酶基因scsamdc的克隆和转化烟草的研究生物工程专业论文.docxVIP

(5)成功构建含有ScSAMDC3基因的CaMV35S组成型启动子的表达 (5)成功构建含有ScSAMDC3基因的CaMV35S组成型启动子的表达 载体pBll21．SeSAMDC3，并通过农杆菌介导法，将ScSAMDC3基因整合入 烟草基因组中。为后一步烟草中过量表达ScSAMDC3基因，增强转基因烟 草抗旱性研究打下基础。 本研究进一步阐明了S一腺苷蛋氨酸脱羧酶基因参与甘蔗抗旱胁迫的分 子机制，丰富了作物抗旱基因的基因库，为其他作物抗旱遗传改良提供候 选基因。 关键词：甘蔗；S．腺苷蛋氨酸脱羧酶；克隆；聚乙二醇；表达分析；基因； 遗传转化 Il 万方数据 RESEARCHS RESEARCHS oN MoLECULAR CLoNING oF SUGARCANE S．ADENoSYLMETHIoNINE DECARBoXYLASE GENE AND ITS GENETIC TRANSFoRMATION TO TABACCO ABSTRACT In this study,the mORF of S-adenosylmethionine decarboxylase gene from sugarcane(Saccharum officinarum)was cloned．Sequence characteristics， biological information of coding regions and expression characteristic analysis was also conducted．It lays the foundation for the research of gene genetic transformation．The study mainly achieved the following results： (1)ScSAMDCl，ScSAMDC2 and ScSAMDC3 gene mORF from sugarcane was cloned by homologous cloning．The mORF of ScSAMDCl，whose length was 1 1 8 8 bp，encoded 3 95 amino acid residues，and its predicted protein molecular weight was 42．78 kD．The mORF of ScSAMDC2,whose length was 1 200 bp，encoded 399 amino acid residues，and its predicted protein molecular weight was 43．53kD．The mORF of ScSAMDC3，whose length was 1 1 88 bp， encoded 395 amino acid residues，and its predicted protein molecular weight was 42．96 kD． (2)Amino acid sequence analysis indicated that，three enzymes were zymogen hydrophilic protein，and the protein structure were not stable． Moreover,the three zymogens contained two highly conserved functional domains—PEST domains and proenzyme cleavage site． (3)Phylogenetic tree analysis showed that，the amino acid sequence deduced from the ScSAMDC gene，which were more closed to gramineous plants 1ike com，millet，sorghum，spot spear and wheat，was in a branch of monocotyledons．Howeveg the three SAMDC of sugarcane were not clustered into the same small branch． 111 万方数据 (4)Realtime (4)Realtime fluorescence quantitative PCR analysis showed that，the expression of ScSAMDC2 and ScSAMDC3 were strongly induced by