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根据其它植物 ACC合成酶 ( 1 - aminocyclopropane- 1 - carboxylic acid synthase,ACS)氨基酸保守区 ,设计 1组简并引物 ,用 RT- PCR法 ,从柿 ( Diospyros kaki Thunb.)果实扩增出3个约 1 kb左右的 c DNA片段 ,将其克隆至 p GEM- T载体上 ,对这些重组克隆进行序列测定和氨基酸序列推导 ,DK- ACS1由 1 1 0 1个碱基组成 ,编码 364个氨基酸 ;DK- ACS2是 1 0 86个碱基 ,编码 35 9个氨基酸 ;DK- ACS3为 1 0 89个碱基 ,编码 363个氨基酸。它们均具有其它植物 ACS合成酶中存在的 7个保守区和 1 1个不变氨基酸残基 ,且在多肽水平上有较高的同源性。与番茄 L E- ACS2的同源性 DK- ACS1是 60 .5 % ,DK- ACS2是 70 .7% ,DK- ACS3为66.9% ;与甜瓜 CM- ACS1的同源性依次分别是 60 .4% ,72 .1 %和 64.4%。
A set of degenerate primers was designed based on the amino acid conserved regions of other plant ACC synthase (1 - aminocyclopropane - 1 - carboxylic acid synthase, ACS), and 3 primers were amplified from the fruit of Diospyros kaki Thunb. About 1 kb of c DNA fragment was cloned into p GEM-T vector, and the recombinant clones were sequenced and deduced amino acid sequence. DK-ACS1 consisted of 1110 bases and encoded 364 amino acids. DK-ACS2 is 1 086 bases, encoding 359 amino acids; DK-ACS3 is 1 089 bases, encoding 363 amino acids. All of them have seven conserved regions and 11 unmodified amino acid residues in other plant ACS synthetases, and have high homology at the polypeptide level. The homology to tomato L-ACS2 was 60.5% for DK-ACS1, 70.7% for DK-ACS2 and 66.9% for DK-ACS3. The homologies to melon CM-ACS1 were 60.4 %, 72.1% and 64.4%.