【摘要】 根据番茄１－氨基环丙烷羧酸合成酶（ＡＣＣ合成酶）基因ｃＤＮＡ序列，按照锤头状核酶作用模式，设计合成长度分别为４０ｍｅｒ和４８ｍｅｒ的一对引物，经体外退火、延伸、连接，插入质粒ｐＧＥＭ－３Ｚｆ（＋）的ＫｐｎＩ和ＢａｍＨＩ位点之间，经菌落原位杂交、双酶切、聚丙烯酰胺凝胶电泳和序列分析证明，人工合成了特异切割ＡＣＣ合成酶ｍＲＮＡ第６６７～６６９位点ＧＵＣ序列的核酶基因序列。更多还原

【Abstract】 In order to interrupting the ACC synthase activity in tomato, a hammerhead ribozyme targeting to cleave GUC triplet at the position 667～699 of ACC synthase mRNA was established and integrated into Kpn I and Bam HI site of vector pGEM- 3Zf(+). Four positive clones were gotten by in situ hybridization. The recombinatant plasmids were digested with restriction endonuclease Kpn I and Bam HI. By polyacrylamide gels electrophoresis and sequence analysis, the size and the sequence of one of the digested product was the same as those of what we expected. These results showed that the ribozyme gene sequence has been obtained.更多还原