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通过对大豆耐盐品种文丰7盐处理抑制差减文库的筛选,获得了一些差异表达的EST序列,与NCBI中EST数据库进行比对分析后发现,其中1个与硫氧还蛋白相关。据此预测了大豆硫氧还蛋白(Thioredoxin,Trx)基因的cDNA序列,并采用RT-PCR方法克隆了大豆Trx基因。生物信息学分析表明:该基因包含1个354 bp的完整开放阅读框,编码118个氨基酸,与拟南芥和烟草的同源性分别为72%和76%。Real-time PCR结果显示:盐处理后,Trx基因在耐盐和盐敏感品种中的表达量均有所提高,但耐盐品种的上调幅度显著高于盐敏感品种,说明其可能具有提高大豆品种耐盐性的作用。
Some differential expression EST sequences were obtained through the screening of the subtractive library of soybean salt tolerant Wenfeng 7 salt, and compared with the EST database of NCBI, one of them was found to be related to thioredoxin. Based on this, the cDNA sequence of soybean thioredoxin (Trx) gene was predicted and the soybean Trx gene was cloned by RT-PCR. Bioinformatics analysis showed that the gene contains a complete open reading frame of 354 bp, encoding a polypeptide of 118 amino acids with 72% and 76% homology with Arabidopsis and tobacco, respectively. The results of Real-time PCR showed that the expression level of Trx gene in salt-tolerant and salt-sensitive varieties increased after salt treatment, but the up-regulation rate of salt-tolerant varieties was significantly higher than that of salt-sensitive varieties, Salt tolerance effect.