超级稻DAD1基因的cDNA克隆及其正反义表达载体的构建

Molecular Cloning of Super Hybrid Rice DAD1 and Construction of Plant Expression Vector for Sense and Anti-Sense

采用RT-PCR方法从超级稻幼苗中扩增DAD1基因的cDNA,并克隆测序,核酸序列表明该基因编码区为342bp,编码114个氨基酸残基.与GenBank发表的序列(XM_472334)相比,有3处碱基不同,分别为105bp处的C(GenBank的为T),148bp处的A(G),149bp处的C(T).将该基因分别正向和反向插入CaMV35S启动子后,构建了基因在植物中的正义、反义表达载体.

DAD1 cDNA from super hybrid rice （Oryza sativa L.） was amplified by RT-PCR, and the coding sequence of the cDNA was analyzed. The result demonstrated that the coding region of DAD1 cDNA is 342 bp, encodes a protein consisting of 114 amino acids. Compared with the sequence in GenBank, there are three different bases from the gene XM_472334. In order to study the function of this gene, binary vectors were constructed, in which the coding region of the gene was placed under the 35S promoter in either sense or antisense orientation.