机构地区: 中国热带农业科学院热带生物技术研究所热带作物生物技术国家重点实验室
出 处: 《生命科学研究》 1999年第4期339-343,共5页
摘 要: 橡胶延伸因子REF(Rubber Elongation Factor)是橡胶生物合成中最重要的酶之一.作者利用REF基因序列设计并合成引物. 通过提取胶乳总RNA 用RT法合成cDNA 后,通过PCR技术扩增并克隆了REF基因和3端非编码区. 序列测定结果表明,REF基因全长414 个碱基,编码138 个氨基酸,3端非编码区长112 bp. 与Goyvaerts E 等人发表的序列比较:REF基因同源率为100% ,3端非编码区有2 bp 的差异,同源率为98% . 将所克隆的REF基因及3非编码区进行地高辛标记, 对胶乳和叶片总RNA 进行点杂交分析,结果表明,胶乳总RNA 呈阳性反应,叶片总RNA 则呈阴性反应. The cDNA of rubber elongation factor gene and its 3non coding region was synthesized by PCR and sequenced. The REF structural gene consists of 414 bp encoding 138 amino acids with a 3 non coding region of 112 bp. Both the nucleotide sequences and the amino acid sequences of the coding region have 100% homology with the REF gene sequences reported by Goyvaerts E et al . The nucleotide sequence of the 3 non coding region have 98% homology with the latter. Dot blot hybridization analysis of latex and leaf total RNA reveal that REF gene is a laticifer specific gene.