桃果实ACC合酶cDNA的克隆

根据其它植物ＡＣＣ合酶氨基酸保守区设计两组简并引物，用套式ＰＣＲ技术从桃成熟果实ｃＤＮＡ中扩增出１．１ｋｂ大小特异性片段，将其克隆至ｐＧＥＭ－Ｔ载体上，对重组克隆ｐＰＡＣＳＩ进行全序列测定表明，插入片段全长１１００个碱基，编码３６６个氨基酸，该基因与番茄、笋瓜、小西葫芦、康乃馨、苹果ＡＣＣ合酶ｃＤＮＡ氨基酸序列同源性分别为７２．７％，７１．３％，７１．１％，６９．１％，６５．４％。ＲＮＡ点杂交表明ｐＰＡＣＳＩ基因随着桃果实成熟表达活性增强，乙烯处理能诱导桃果实该基因的表达。

Cloning of an 1-aminocyclopropane-1-carboxylate cDNA Synthase from Peach Fruit

Synthetic oligonucleotides based on the conversed sequence of 1 aminocyclopropane 1 carboxylate(ACC)synthase from other plants were used to prime the synthesis and amplification of about 1.1kb specific fragment by polymerase chain reaction(PCR)from ripening peach fruit.PCR product was cloned into pGEM T vector.and several recombinant clones were isolated.One of recombinants.pPACSl,was sequenced.The inserted fragment contained l 100bp with an open reading frame of 366 amino acids.The predicted protein from peach ACC synthase cDNA were 72.7%, 71.3%, 71.1%,69.1%,65.4% identical to the deduced proteins from tomato,squash,zucchini,carnation,apple,respectively.pPACSl mRNA accumulated during peach fruit ripening and after ethylene treatment.