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以柿 (DiospyroskakiThunb .)果实为材料 ,根据其它植物ACC氧化酶 (1 aminocyclopropane 1 carboxylicacidoxidase ,ACO)氨基酸保守区 ,设计 1组简并引物 ,通过RT PCR法扩增出 2个约 80 0bp大小的cDNA片段。将其克隆至pGEM T载体上 ,对这 2个重组克隆进行序列测定 ,DK ACO1由 834个碱基组成 ,编码 2 5 9个氨基酸 ;DK ACO2为836个碱基 ,编码 2 6 0个氨基酸。它们均具有其它植物ACC氧化酶中存在的保守区 ,且在多肽水平上的同源性很高 ,与番茄LE ACO1的同源性DK ACO1是 86 .2 % ,DK ACO2是 82 .5 % ;与甜瓜CM ACO1的同源性DK ACO1是82 .6 % ,DK ACO2为 81.1%。
A set of degenerate primers was designed based on the amino acid conserved regions of other plant 1-aminocyclopropane 1 carboxylic acid oxidase (ACO) from the fruit of Diospyroskaki Thunb. Two cDNAs of about 80 0 bp in size were amplified by RT-PCR Fragment. The clones were cloned into pGEM T vector, and the two recombinant clones were sequenced. DK ACO1 consists of 834 bases and encodes a protein of 259 amino acids. DK ACO2 is 836 bases and encodes 260 amino acids. All of them have the conserved regions in ACC oxidase of other plants, and have high homology at the polypeptide level. The homology of DK ACO1 to the tomato LE ACO1 is 86.2% and the DK ACO2 is 82.5%. The homology to melon CM ACO1 was 82.6% for DK ACO1 and 81.1% for DK ACO2.