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结合表达谱数据分析,参考公共数据库中的序列信息设计引物,利用RT-PCR技术,从东乡野生稻中获得了1个在低温诱导条件下高表达的螺旋—环—螺旋结构域蛋白基因BGIOSGA013293-DX的全长c DNA,并构建了其过表达载体。序列比对分析表明,该基因位点在93-11中编码308个氨基酸,而在东乡野生稻中编码335个氨基酸,比93-11多27个氨基酸。且这27个氨基酸连续分布在一起,另外还有1个氨基酸的差异。利用农杆菌介导法将BGIOSGA013293-DX转入水稻受体品种93-11,获得了9株过表达转基因水稻植株,经潮霉素抗性标记基因PCR阳性检测和GUS检测,获得的转基因植株为携带BGIOSGA013293-DX的阳性植株。
Based on the analysis of the expression profile data, primers were designed with reference to the sequence information in the public database and a high-expression helix-loop-helix domain protein gene, BGIOSGA013293-, was obtained from Dongxiang wild rice by RT-PCR. DX full-length c DNA, and constructed its overexpression vector. Sequence alignment analysis showed that this gene encodes 308 amino acids in 93-11, whereas it encodes 335 amino acids in Dongxiang wild rice, 27 amino acids more than 93-11. And these 27 amino acids are continuously distributed together, in addition there is a difference of 1 amino acid. Agrobacterium-mediated transformation of BGIOSGA013293-DX into rice receptor variety 93-11, obtained nine overexpression transgenic rice plants, hygromycin resistance marker gene PCR positive detection and GUS detection, the obtained transgenic plants were Positive plants carrying BGIOSGA013293-DX.