菠菜甜菜碱醛脱氢酶基因的克隆和序列分析  

A cDNA Cloning and Sequence Encoding Betaine Aldehyde Dehydrogenase (BADH) from Spinach

作　　者：马德钦[1] 吕文[1] 汤岚[2] 骆爱玲[2] 梁峥[2] 

机构地区：[1]中国科学院微生物研究所,北京100080 [2]中国科学院植物研究所,北京100044

出　　处：《生物工程学报》1996年第1期65-70,共6页Chinese Journal of Biotechnology

基　　金：国家自然科学基金

摘　　要：以耐盐的菠菜mRNA为模板,经反转录合成甜菜碱醛脱氢酶(BADH)基因第一链cDNA。在人工合成的两端引物引导下,通过多聚酶链式反应(PCR),扩增获得双链cDNA。把重组有BADH基因的pUC19转化至E.coli DH5α菌株,亚克隆后测定了基因的全序列。所得到的BADH基因全长序列为1491bp,编码497个氨基酸。与文献报道的相比较,核苷酸序列同源性99.8％,氨基酸序列同源性达99.6％。在此基础上,构建了BADH基因的高等植物表达载体。A cDNA clone encoding betaine aldehyde dehydrogenase (BADH) has been amplified from spinach mRNA through polymerase chain reaction (PCR) technique using two primers specific to spinach BADH cDNA. The authenticity of the BADH gene was confirmed by cleavage it with restriction endonuclease and nucleotide sequence analysis. This analysis demonstrated the presence of a 1491 bp open reading frame and of restriction cleavage sites identical with the previously determined sequence. Homology of the nucleotide sequence and the deduced amino acid sequence in the spinach BADH gene with the previously determined sequence show 99- 8% and 99. 6% respectively. A plant expresion vector with BADH gene has been constructed.

关 键 词：序列分析 菠菜 甜菜碱 醛脱氢酶 基因克隆 

分 类 号：S636.103.2[农业科学—蔬菜学]