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将编码水稻非特异性脂质转移蛋白 (nonspecificlipidtransferprotein ,nsLTP)基因 (LTP110 )的克隆到硫氧还蛋白融合表达载体PET32a(+)中 ,在BL2 1(DE3)trxB-宿主菌中实现了融合蛋白的高表达。通过Ni2 + chelatingSepharosefastflow柱纯化融合蛋白后 ,通过肠激酶酶切再过该亲和柱得到了重组LTP110。CD谱扫描表明重组蛋白质与体内提取的nsLTP二级结构相似 ;荧光脂质结合实验表明该蛋白质具有结合脂肪酸分子的活性。对该蛋白质的抑菌功能进行研究后表明 ,LTP110具有抑制稻瘟病菌孢子萌发的功能 ,在较低浓度即能发挥活性
The nonspecific lipid transferrin (nsLTP) gene (LTP110) was cloned into the thioredoxin fusion expression vector PET32a (+) and the fusion protein was obtained in BL21 (DE3) trxB- High expression. After purification of the fusion protein by Ni2 + chelating Sepharose fast flow column, recombinant LTP110 was obtained by digestion with enterokinase and then through the affinity column. CD spectra showed that the recombinant protein was similar to the secondary structure of nsLTP extracted in vivo. Fluorescence lipid binding assay showed that the protein possessed the activity of binding fatty acid molecules. The study of the bacteriostatic activity of the protein showed that LTP110 has the function of inhibiting the spore germination of Magnaporthe grisea and can exert its activity at a lower concentration