[Objectives]This study was conducted to construct a pColdI-HSP70 recombinant prokaryotic expression vector.[Methods]With rice‘N22’as the test variety,hydroponic experiments were set up for rice seedlings with normal...[Objectives]This study was conducted to construct a pColdI-HSP70 recombinant prokaryotic expression vector.[Methods]With rice‘N22’as the test variety,hydroponic experiments were set up for rice seedlings with normal growth(CK),high temperature treatment(H),drought treatment(D)and drought-high temperature cross treatment(DH).The recombinant prokaryotic expression vector was constructed by the method of prokaryotic expression,and its induction expression time,IPTG concentration and temperature were optimized.[Results]The pColdI-HSP70 expression vector was successfully constructed,and the fusion protein was highly expressed in host strain BL21,and the expressed proteins were all in a soluble form.By optimizing the induction expression conditions,it was found that the optimal expression conditions were the IPTG concentration of 0.5 mmol/L and induction at 20℃for 36 h.The expression analysis of the rice HSP70 gene under different stress treatments was carried out by qRT-PCR technology,and it was found that H,D and DH stresses all could induce its expression,and its expression levels were 4.65,1.40 and 17.66 times higher than that of the CK group,respectively.[Conclusions]This study lays a solid foundation for the isolation,purification and functional study of rice HSP70 proteins.展开更多
Tumor necrosis factor receptor(TNFR)-associated factor 6(TRAF6)is a key regulator of the innate immune system,activating NF-κB and AP-1 signaling[1].TRAF6 participates in the immune regulation of many diseases,especi...Tumor necrosis factor receptor(TNFR)-associated factor 6(TRAF6)is a key regulator of the innate immune system,activating NF-κB and AP-1 signaling[1].TRAF6 participates in the immune regulation of many diseases,especially in chronic infections caused by pathogenic bacteria,such as Mycobacterium tuberculosis(Mtb)-induced tuberculosis(TB)[2].展开更多
基金Supported by Anhui Provincial Education Department Project(KJ2019A0213)National Natural Science Foundation of China(31501245)Undergraduate Innovation and Enterpreneurship Training Program of Anhui Province(S202010364185,XJDC2020541,202110364788)。
文摘[Objectives]This study was conducted to construct a pColdI-HSP70 recombinant prokaryotic expression vector.[Methods]With rice‘N22’as the test variety,hydroponic experiments were set up for rice seedlings with normal growth(CK),high temperature treatment(H),drought treatment(D)and drought-high temperature cross treatment(DH).The recombinant prokaryotic expression vector was constructed by the method of prokaryotic expression,and its induction expression time,IPTG concentration and temperature were optimized.[Results]The pColdI-HSP70 expression vector was successfully constructed,and the fusion protein was highly expressed in host strain BL21,and the expressed proteins were all in a soluble form.By optimizing the induction expression conditions,it was found that the optimal expression conditions were the IPTG concentration of 0.5 mmol/L and induction at 20℃for 36 h.The expression analysis of the rice HSP70 gene under different stress treatments was carried out by qRT-PCR technology,and it was found that H,D and DH stresses all could induce its expression,and its expression levels were 4.65,1.40 and 17.66 times higher than that of the CK group,respectively.[Conclusions]This study lays a solid foundation for the isolation,purification and functional study of rice HSP70 proteins.
基金supported by grants from the National Key R&D Program of China(2023YFC2307301,2022YFC2302900 and 2021YFA1300902)Chinese National Program on Key Basic Research Project(2017YFA0505900)+1 种基金National Natural Science Foundation of China(9184230036,31730025,91542111,81330069,31030028,30525012,82122029,82102400and 82071776)the fellowship of China National Postdoctoral Program for Innovative Talents(Bx2021215).
文摘Tumor necrosis factor receptor(TNFR)-associated factor 6(TRAF6)is a key regulator of the innate immune system,activating NF-κB and AP-1 signaling[1].TRAF6 participates in the immune regulation of many diseases,especially in chronic infections caused by pathogenic bacteria,such as Mycobacterium tuberculosis(Mtb)-induced tuberculosis(TB)[2].
