番茄1位氨基环丙烷羧酸(ACC)合成酶基因的分离和克隆

Isolation and Cloning of the Gene Encoding1-aminocyclopropane-1-Carboxylate Synthase in Tomato

以番茄基因组ＤＮＡ为模板，利用多聚酶链反应技术（ＰＣＲ）将ＬＥ－ＡＣＣ２基因的编码区的第四外显子进行扩增．所得的ＤＮＡ扩增片段克隆至质粒ｐＥＧＭ－３ｚｆ（＋）的ＢａｍＨＩ和ＨｉｎｄⅢ位点之间，并用限制内切酶酶切、ＰＣＲ扩增及ＤＮＡ序列分析等手段证实已获得ＬＥ－ＡＣＣ２第四外显子序列的克隆．该克隆的碱基序列与国外报道的相关序列相一致，本文还对该基因的反义片段在番茄延熟品种培育上的意义进行了阐述和探讨．

Using tomato genomic DNA as template, the fourth exon's region of LE-ACC2 gene was amplified by Polymerase Chain Reaction (PCR). The DNA fragment obtained from PCR was cloned the site between BamHⅠ and HindⅢ in the vector pGEM-3zf(+). The recombinant plasmid was screened under digestion of restriction endonuclease, amplification of Polymerase Chain Reaction and DNA sequencing, the results showed that the base sequences of the fourth exons between ours and others are fully uniform. The value of the antisense fragments of the ACC gene to delaying fruit ripening is also described and discussed.