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为了进一步研究小麦谷胱甘肽-S-转移酶基因(TaGST)的功能,采用RT-PCR方法分离了小麦谷胱甘肽-S-转移酶基因(TaGST)的ORF全长c DNA,并进行了生物信息学分析。结果表明:小麦TaGST基因的ORF全长690 bp,编码229个氨基酸;TaGST蛋白分子质量为25.81 k Da,p I为5.29。系统进化分析表明,该基因编码蛋白与水稻OsGST蛋白的氨基酸同源性最高,与已知植物GST家族成员的氨基酸序列聚类分析将TaGST聚为Phi类GST。构建原核表达载体p ET32-TaGST,对TaGST基因进行原核表达,SDS-PAGE结果表明,其所表达蛋白与预期蛋白大小一致。为进一步研究该基因的特性和功能奠定了理论基础。
To further investigate the function of wheat glutathione-S-transferase gene (TaGST), ORF full length cDNA of wheat glutathione-S-transferase gene (TaGST) was isolated by RT-PCR and performed Bioinformatics analysis. The results showed that the ORF of TaGST gene in wheat was 690 bp in length and encoded 229 amino acids. The molecular weight of TaGST protein was 25.81 kDa, and pI was 5.29. Phylogenetic analysis showed that the deduced amino acid sequence of this gene was the highest homology with rice OsGST protein. According to the amino acid sequence analysis of GST family members, TaGST was clustered into Phi GST. The prokaryotic expression vector p ET32-TaGST was constructed and the prokaryotic expression of TaGST gene was carried out. SDS-PAGE results showed that the expressed protein was consistent with the expected protein size. Which laid a theoretical foundation for further study on the characteristics and functions of this gene.