【作者】 韩蕾； 和绍禹； 董捷； 黄家兴； 吴杰；

【Author】 Han Lei;He Shaoyu;Dong Jie;Huang Jiaixng;WU Jie;Eastern Bee Research Institute,Yunnan Agricultural University;Key Laboratory for Insect-Pollinator Biology of the Ministry of Agriculture,Institute of Apicultural Research,Chinese Academy of Agricultural Sciences;

【机构】 云南农业大学东方蜜蜂研究所； 中国农业科学院蜜蜂研究所农业部授粉昆虫生物学重点开放实验室；

【摘要】 兰州熊蜂是我国优良熊蜂蜂种之一,可用于大棚温室授粉。鞘磷脂磷酸二酯酶(SMPD)是一种水解酶,参与鞘磷脂代谢过程,并起关键作用。本研究从诱导温度、IPTG浓度及诱导时间三个方面进行优化,以获得最佳原核表达条件。结果表明:诱导温度与诱导时间是影响SMPD蛋白表达量的主要因素,而诱导剂IPTG浓度影响较小。当温度为10℃,IPTG终浓度为0.1 mM,诱导时间为32 h时,目的蛋白表达量较高,特异性较好,能够满足该蛋白后续纯化实验要求,为进一步开展SMPD蛋白功能研究奠定基础。更多还原

【Abstract】 Bomhus lantschouensis is an excellent pollinating species for greenhouse crop pollination.To improve the expression level of the SMPD protein in E.coli BL21(DE3),the prokaryotic expression conditions were optimized from the inducement temperature,concentration of IPTG and inducement time.The result indicat that inducement temperature and time was the main factor for the expression level of the SMPD protein,while the concentration of IPTG had minor effect.The highest expression level of the target protein is as follow:inducement temperature at 10 ℃,the concentration of 0.1 mM of the IPTG and 32 h inducement time.The current study provides a basis for studying the role of SMPD protein in Bombus lantschouensis.更多还原