用酵母双杂交系统研究载脂蛋白AI(apoAI)和清道夫受体BI(SR-BI)间的相互作用  

Study on Interaction Between Apolipoprotein AI and Scavenger Receptors Class B Type I by Yeast Two-Hybrid

作　　者：信学雷[1] 陈志慧[1] 李维琪[1] 刘云英[1] 麦迪娜[1] 张云峰[1] 

机构地区：[1]中国科学院新疆理化技术研究所,乌鲁木齐830011

出　　处：《中国生物工程杂志》2005年第5期80-84,共5页China Biotechnology

基　　金：中国科学院"西部之光"资助项目

摘　　要：目的:利用酵母双杂交系统验证在胆固醇逆转运过程中起关键作用的大鼠载脂蛋白AI(apoAI)和清道夫受体BI- (SR -BI)间存在着相互作用,为初步筛选具有降脂活性组分提供1对新的靶点。方法:首先分别克隆了Wistar大鼠的apoAI和SR -BI基因的cDNA ,并构建了酵母表达载体,利用共转化技术观察到apoAI和SR -BI间存在着相互作用,并在酵母交配实验中证实了这个结果。结果:经共转化后的实验组与阳性对照组可在SD/ -Leu/- Trp/- His/- Ade/X -α- Gal平板上生长且菌斑呈蓝色,经测定α、β半乳糖苷酶活力可知酶活分别为8～1 2U和1 0～40U。酵母交配后的二倍体实验组、阳性对照组可在SD/- Leu/- Trp/ -His/- Ade/X -α- Gal平板上生长且菌斑呈蓝色。结论:apoAI和SR BI间的确存在相互作用。Objective: To verify the interaction between apolipoprotein AI and scavenger receptors class B type I(SR|BI), a key process of reverse cholesterol transport, by yeast two-hybrid method and to provide a new target for selecting active substance for reducing lipid in plasma. Method: Apolipoprotein AI and SR|BI gene cDNA from Wistar rat were cloned, and DNA/apoAI and AD/SR-BI were constructed, then co-transformed into yeast strain AH109. Transformants were selected on medium of SD/-Leu/-Trp/-His/-Ade/X-α-Gal and the putative positive clones were identified by yeast mating. Result: Transformants can grow on medium of SD/-Leu/-Trp/-His/-Ade/X-α-Gal and the colonies are blue.α?β- galactosidase assays show that the activity of α?β-galactosidase are 8～12U and 10～40U respectively. After yeast mating, blue colonies can still be seen on medium of SD/-Leu/-Trp/-His/-Ade/X-α-Gal. Conclusion: It is true that there is interaction between apolipoprotein AI and SR|BI.

关 键 词：酵母双杂交系统 载脂蛋白AI 清道夫受体BI 相互作用 胆固醇 逆转运 降血脂药物 基因表达 

分 类 号：Q786[生物学—分子生物学] R972.6[医药卫生—药品]