摘要： 利用3′RACE的方法对柿果实ACC合成酶基因的3′末端进行扩增，扩增产物克隆到pMD18-T Vector载体上，转化到大肠杆菌感受态细胞DH5α，蓝白斑筛选重组质粒，并对插入片段进行序列测定。结果表明，3′RACE产物长912 bp，开放阅读框内核苷酸序列编码251个氨基酸，此氨基酸序列含有ACC合成酶中3个高度保守区，GenBank中登录的DK-ACS1（登录号AB073005）序列只有一个的氨基酸不同。终止子后面的3′非编码区的长度为155 bp，其中只有一个核苷酸与DK-ACS1的3′序列非编码区不同。序列分析结果显示，克隆的片段含有5′端GSP Inner Primer和3′端为RACE Inner Primer的引物。

关键词: 柿果, 富有, ACC合成酶, 3&prime, RACE

Abstract: Gene cloning is the bases to research gene function and expressing characterization.The popular used method is to partial clone gene coding sequence first by the gene specific prime desired from the gene conserve amino acid,then to get 5'and 3'end sequence which is much difficult than coding region cloning.A cDNA fragment with a 3'end sequence of ACC synthase was cloned from Fuyu persimmon fruit by 3'RACE PCR.The PCR product was linked into the pMD18-T Vector and recombinant plasmid was transformed to E.coli DH5α,which was selected with xgal/IPTG.Sequence analysis showed the PCR product was 912 bp,which encoded 251 amino acids with three highly conserved regions of ACC synthase.There was only one amino acid different from the sequence DK-ACS1 which entered in GenBank(AB073005).There was a 155 bp non-encoding sequence in 3'end,in which only one nucleotide was different from DK-ACS1.The sequence included 5'GSP inner primer and 3'RACE inner primer,which indicated to get a full 3'end sequence of Fuyu persimmon fruit.

Key words: Persimmon, Fuyu, ACC synthase, 3’RACE

中图分类号: 

• S665.2