Molecular characteristics, recombinant expression and activity detection of OsGSTL1 from rice

Brazilian Archives of Biology and Technology

Tingzhang Hu²  Yongwei Yang²  Lili Tan²  Junnian Yang¹  Yingmei Wu¹ 

[1] ,Chongqing University College of Bioengineering Key Laboratory of Biorheological Science and Technology,China

关键词: glutathione S-transferase;    GST activity;    In silico;    Oryza sativa L.;    recombinant expression;    semi-quantitative RT-PCR;   

DOI  :  10.1590/S1516-89132014000100011

来源: SciELO

【 摘 要 】

The mRNA of OsGSTL1 was detected in the roots and leaves of rice plants at seedling and tillering stages, and their roots, leaves and panicles at the heading stage. The full-length open reading frame of OsGSTL1 cDNA was 732 bp and encoded a putative polypeptide of 243 amino acids with a calculated molecular mass of 27.30 kDa and a theoretical pI of 5.50. The protein sequences of OsGSTL1 exhibited typical feature of the lambda class GST, which contained the conserved domain "GST_C_Lambda" in C-terminal alpha helical domain and a highly conserved Cys42 in active center. In silico predictions showed that the OsGSTL1 protein was strongly hydrophilic. The phylogenetic analysis revealed OsGSTL1 belonged to monocots subgroup and was closer to IN2-1 of Z. may. The OsGSTL1 gene was cloned into pYTV vector and was introduced into yeast strain PEP4. Western blot analysis showed that the exogenous OsGSTL1 was expressed in the transformed yeast. The GST activity of the crude extracts of yeast showed that the OsGSTL1 transgenic yeast had higher levels of GST activities than the control yeasts. These findings suggested that the OsGSTL1 was a glutathione S-transferase and could play an important role during the growth and development processes in rice.

【 授权许可】

CC BY-NC   
 All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License