酸橙ACC氧化酶基因片段克隆及序列分析

1-氨基环丙烷-1-羧酸(ACC)氧化酶是植物乙烯合成的1个关键酶,乙烯作为一种内源激素,对植物生长、老熟过程有多方面的调节作用。根据已报道的各种植物ACC氧化酶氨基酸序列上前后两个保守区设计1对简并引物,以酸橙叶片总RNA为模板,通过RT-PCR扩增到1个834 bp的基因片段。克隆了酸橙1-氨基环丙烷-1-羧酸(ACC)氧化酶基因cDNA片段,将片段序列在NCB I网站上进行同源性搜索,显示的皆为不同植物的ACC氧化酶基因,因而认为所克隆的片段就是酸橙ACC氧化酶基因;并运用DNAStar5.0软件进行序列分析,推导的氨基酸序列为278个残基;具有所有植物ACC氧化酶基因共有的保守区域;与多种植物的ACC氧化酶基因的核苷酸和氨基酸序列的同源性都在72%和70%以上。

ACC Oxidase Gene Cloning and Sequence Analysis of Citrus aurantium L.

1-aminocyclopropane-1-carboxylate(ACC) oxidase is a crucial enzyme in ethylene synthesis in plant.The plant hormone ethylene is not only responsible for the initiation of fruit ripening and senescence but also for regulating many other plant developmental processes.To establish the base of the control of the Citrus aurantium L.fruit maturity in molecular level and produce high quality Fructus Aurantii,a pair of degenergte primers were designed according to the conserve regions of ACC Oxidase gene of many kinds of plants.The cDNA document of ACC oxidase gene of Citrus aurantium L. was cloned.By using the program of BLAST on NCBI GenBank database,the sequence presented a very high match with the ACC oxidase genes from other plants.The base sequence was analysed by using biology programe of DNAStar 5.0.278 amino acids were coded by the base sequence and the base sequence had the same conserve region of ACC oxidase gene of many kinds of other plants.The base sequences comparability was more than 72% compared with those of many kinds of other plants. The amino acid sequences comparability was more than 70% compared with those of a lot of other plants.