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采用RT-PCR方法从小麦品种豫教2号的发育籽粒中克隆出小麦淀粉合成关键酶AGPase胞质型小亚基(AGPase plastic small subunit,SSU I)的cDNA全长,并构建了此基因的过表达、反义表达和RNAi干扰载体.SSUI的cDNA全长为1465 bp(GenBank No.EF405961),编码域长度为1 422 bp,位于EF405961的2～1 423 bp.同源性比较结果表明:与GenBank上已报道的SSU I基因(X66080,AF244 997,Z48562)同源性达98%～99%.以pWM101质粒为基础,构建了由35 S启动子调控的SSU I基因的过表达载体pWM101SSU I S;将该基因反向置于pBI121质粒的CaMV35S启动子之后,构建了SSU I的反义表达载体pBI121SSU I A.同时还克隆出SSU I基因的一段260 bp高保守序列,以pFGC5941质粒为基础,构建了RNAi干扰载体pFGC5941SSU I.
The full-length cDNA of AGPase plastic small subunit (SSU I), a key enzyme in the synthesis of wheat starch, was cloned from the developmental kernels of wheat cultivar Yujiao 2 by RT-PCR. Overexpression, antisense expression and RNAi interference vectors.The full-length cDNA of SSUI was 1465 bp (GenBank No.EF405961), with a coding region of 1 422 bp located between 2 and 4243 bp of EF405961.The homology comparison showed that: Which shared 98% -99% homology with the reported SSU I gene (X66080, AF244 997, Z48562) in GenBank.Overexpression vector pWM101SSU of SSU I gene regulated by 35S promoter was constructed based on pWM101 plasmid IS, the antisense expression vector pBI121SSU I A of SSU I was constructed after reverse transposing this gene into the CaMV35S promoter of pBI121 plasmid. A 260 bp highly conserved sequence of the SSU I gene was also cloned and was based on the pFGC5941 plasmid , Constructed the RNAi interference vector pFGC5941SSU I.