豆科凝集素基因Le4的克隆及其表达产物对蚜虫的抗性

发布时间：2018-06-05 10:58

本文选题：植物凝集素 + Le　；参考：《基因组学与应用生物学》2016年12期

【摘要】：植物凝集素是含有一个或者多个非催化结构,并能可逆结合特异单糖或寡糖的蛋白,它具有凝集细胞、沉淀多糖和糖蛋白的功能。近年来,植物凝集素因其独特的生理功能,在农作物抗虫工程中获得了广泛的关注。本研究采用RT-PCR的方法,从浙江苞萝黄大豆叶片中成功克隆了一个新的豆科凝集素Le4基因,利用p ET-28a质粒构建了Le4基因的原核表达载体,成功表达了Le4蛋白。通过对诱导温度及异丙基-β-D-硫代半乳糖苷(IPTG)终浓度等条件进行优化,利用Western blotting技术,在Le4表达菌株超声破碎后的上清液中检测到了可溶性的Le4蛋白。通过涂抹法将含有可溶性Le4表达蛋白的大肠杆菌菌液涂抹在小麦叶片上,与涂抹空载体菌液相比,小麦叶片麦长管蚜数目减少了约50%。以上结果初步表明来源于大豆的凝集素基因Le4蛋白具有杀虫效应。
[Abstract]:Plant lectin is a protein with one or more non-catalytic structures that can reversibly bind to specific monosaccharides or oligosaccharides. It has the functions of agglutinating cells, precipitating polysaccharides and glycoproteins. In recent years, plant lectin has gained wide attention in crop insect resistance engineering because of its unique physiological function. In this study, a new legume lectin Le4 gene was successfully cloned from the leaves of Zhejiang Buxiahuang soybean by RT-PCR method. The prokaryotic expression vector of Le4 gene was constructed by using p ET-28a plasmid, and the Le4 protein was successfully expressed. By optimizing the induction temperature and the final concentration of isopropyl- 尾 -Dthioglycoside, soluble Le4 protein was detected in the supernatant of Le4 expression strain after ultrasonic crushing by Western blotting technique. The Escherichia coli solution containing soluble Le4 expression protein was applied to wheat leaves by smearing. Compared with the empty carrier solution, the number of wheat aphid in wheat leaves was reduced by about 50%. These results indicated that the agglutinin gene Le4 protein derived from soybean had insecticidal effect.
【作者单位】：南昌大学生命科学学院江西省分子生物学与基因工程重点实验室;江西省超级水稻研究发展中心;
【基金】：国家自然科学基金(31401038,31360338) 2015年度南昌大学研究生创新专项资金(cx2015103)共同资助
【分类号】：Q943.2;S43

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