Chinese Journal OF Rice Science >
Heterologous Expression and Oligomeric Identification of a Small Heat Shock Protein (SHSP) from Oryza sativa
Received date: 2017-01-18
Revised date: 2017-02-21
Online published: 2017-09-10
【Objective】In our previous study, a small heat shock protein gene OsSHSP17.6 was cloned from Oryza sativa and its expression was shown to be significantly up-regulated by heat shock or viral infection, suggesting that the OsSHSP17.6 could play an important role in both biotic and abiotic stress responses. In this study, our objective is to further identify the characteristics of OsSHSP17.6. 【Method】 The OsSHSP17.6 gene was sub-cloned into the plasmid pET-32a, a prokaryotic expression vector, and transformed into Escherichia coli BL21(DE3)pLysS for inducible expression. Then the recombinant protein was purified with affinity chromatography and used for native PAGE and Western-blotting assays. 【Result】Its heterologous expression appeared to alleviate the poisonous effect of IPTG on the host E. coli. Native PAGE and Western-blotting assays showed that the purified recombinant OsSHSP17.6 could form homological dimers and oligomer in vitro. 【Conclusion】 Taken together, these findings supported the hypothesis that the protein should be functional molecular chaperone in vivo and indicated that OsSHPS17.6 could be involved in the stress response by its homological oligomerization, which could contribute to the functional identification of OsSHSP17.6.
Nianjun CAI, Liuming GUO, Jing LI, Congying XIANG, Jian YANG, Jianping CHEN, Hengmu ZHANG . Heterologous Expression and Oligomeric Identification of a Small Heat Shock Protein (SHSP) from Oryza sativa[J]. Chinese Journal OF Rice Science, 2017 , 31(5) : 483 -488 . DOI: 10.16819/j.1001-7216.2017.7010 483
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