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以卡特兰(Cattleya)花朵为试材,首先提取总RNA,并根据其它兰花的ACC氧化酶(1-aminocyclopropane-1-carboxylic acid oxidase,ACO)基因保守序列设计1对特异性引物,然后通过RT-PCR法,克隆得到一条卡特兰ACC氧化酶的cDNA片断,该片断长度为967 bp,共编码321个氨基酸残基。序列分析结果显示该克隆片断与已发表的其它兰花的ACC氧化酶基因序列同源性很高,均在85%以上,尤其与其近亲C.bicolor、C.inter-media、Laelia anceps的同源性均在95%以上。
Using Cattleya flower as test material, total RNA was extracted first and a pair of specific primers were designed according to the conserved sequence of ACC-oxidase (ACO) in other orchids and then analyzed by RT -PCR method, a cDNA fragment of Cattell ACC oxidase was cloned. The fragment was 967 bp in length and encoded a total of 321 amino acid residues. Sequence analysis showed that the cloned fragment shared high homology with ACC oxidase genes of other orchids, all of which were more than 85%, especially with homologous C.bicolor, C.inter-media and Laelia anceps All above 95%.