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目的分析重组半乳糖苷凝集素AAL抑制烟草花叶病毒(Tobacco mosaic virus,TMV)的侵染活性及与TMV衣壳蛋白(CP)的体外结合活性,为进一步明确AAL的生物学功能提供理论依据。方法将含AAL基因的重组表达质粒pET-22b-aal转化大肠杆菌BL21(DE3),IPTG诱导表达,采用Ni-NTA亲和层析纯化重组AAL。采用半叶法分析重组AAL抑制TMV的侵染活性;凝胶复性覆盖法分析重组AAL与TMV-CP的体外结合活性。结果表达及纯化的重组AAL相对分子质量约为18 000;重组AAL具有与从杨树菇子实体中提取的AAL相近的抑制TMV侵染的活性,并可与TMV-CP体外结合。结论重组AAL具有抗TMV的活性,其机理可能是通过与TMV-CP的互作抑制病毒侵染。
Objective To analyze the inhibitory effect of recombinant Galectin-AAL on the infection activity of Tobacco mosaic virus (TMV) and the in vitro binding activity with TMV capsid protein (CP) to provide a theoretical basis for further clarifying the biological function of AAL . Methods The recombinant plasmid pET-22b-aal containing AAL gene was transformed into E. coli BL21 (DE3) and induced by IPTG. Recombinant AAL was purified by Ni-NTA affinity chromatography. The half-leaf method was used to analyze the recombinant AAL to inhibit the infection activity of TMV. The gel-bound method was used to analyze the in vitro binding activity of recombinant AAL and TMV-CP. Results The relative molecular mass of recombinant AAL expressed and purified was about 18 000. Recombinant AAL had the activity of inhibiting TMV infection similar to that of AAL extracted from poplar mushroom fruiting bodies and could be combined with TMV-CP in vitro. Conclusions Recombinant AAL possesses anti-TMV activity and its mechanism may be that it inhibits viral infection through interaction with TMV-CP.