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.
出处
《生命科学研究》
CAS
CSCD
1999年第4期339-343,共5页
Life Science Research
基金
国家自然科学基金!(397690014)
关键词
橡胶
橡胶延伸因子
克隆
点杂交
rubber(cis 1 4 polyisoprene)
rubber elongation factor
cloning
dot blot hybridization
