Longstaff M, Powell K S, Gatehouse J A, Raemaekers R, Newell C A, Hamilton W D
Axis Genetics, Babraham, Cambridge, UK.
Eur J Biochem. 1998 Feb 15;252(1):59-65. doi: 10.1046/j.1432-1327.1998.2520059.x.
Recombinant snowdrop lectin was produced in Escherichia coli from a cDNA clone encoding mature Galanthus nivalis agglutinin. After induction with isopropylthio-beta-D-galactoside, inclusion bodies from E. coli were solubilised and the G. nivalis agglutinin purified by metal-affinity chromatography using a carboxy-terminal hexahistidine tag. The protein was refolded on the metal-affinity column prior to elution. After purification, the recombinant G. nivalis agglutinin agglutinated rabbit erythrocytes to a dilution similar to that determined for 'native' lectin purified from snowdrop, and showed similar specific binding to mannose. The toxicity of the recombinant G. nivalis agglutinin towards rice brown planthopper (Nilaparvata lugens) was shown to be similar to that of 'native' G. nivalis agglutinin when incorporated into an artificial diet. The recombinant G. nivalis agglutinin is thus functionally similar to 'native' snowdrop lectin.
重组雪花莲凝集素是在大肠杆菌中由编码成熟雪花莲凝集素的cDNA克隆产生的。用异丙基硫代-β-D-半乳糖苷诱导后，将大肠杆菌中的包涵体溶解，并使用羧基末端六组氨酸标签通过金属亲和色谱法纯化雪花莲凝集素。该蛋白在洗脱前在金属亲和柱上复性。纯化后，重组雪花莲凝集素凝集兔红细胞的稀释度与从雪花莲中纯化的“天然”凝集素所确定的稀释度相似，并且显示出与甘露糖相似的特异性结合。当掺入人工饲料中时，重组雪花莲凝集素对稻褐飞虱（Nilaparvata lugens）的毒性与“天然”雪花莲凝集素的毒性相似。因此，重组雪花莲凝集素在功能上与“天然”雪花莲凝集素相似。
