摘要
采用三氯乙酸/丙酮沉淀法分别对白木香茎和叶进行蛋白质提取,利用一、两次水化法分别对所获蛋白进行溶解,并通过研究蛋白得率、比较蛋白浓度及经SDS-PAGE单向电泳所获条带的清晰度,分析白木香茎、叶蛋白制备与SDS-PAGE单向电泳条件的优劣。结果表明:白木香茎、叶蛋白得率分别是73mg·g-1与52mg·g-1;一次水化法得到的茎、叶样品蛋白终浓度分别为3.62μg·μL-1,4.17μg·μL-1,两次水化法所获得茎、叶蛋白终浓度为1.53μg·μL-1,2.12μg·μL-1。两种方法所获蛋白样品的SDS-PAGE条带均较模糊,但后者的单向电泳条带杂质较少,究其原因可能是蛋白制备、电泳条件的某些方面仍需改进。该研究为进一步从分子生物学的角度研究白木香结香机制提供了技术的基础。
TCA/acetone precipitation method was employed to extract the proteins of stem and leaves in Aquilaria sinensis ( Lour) Gilg separately ,and one-lysis and twi-lysis were used to dissolve the protein samples .Through stud-ying the harvest rate of proteins extraction and comparing the protein concentration and the clarity of protein bandings from SDS-PAGE single dimensional electrophoresis ,the qualities of the protein preparation and the SDS-PAGE condi-tions were analyzed.The result indicated that,the yield of proteins extraction from stem and leaves were 73mg· g-1 and 52 mg· g-1 respectively ,the protein concentration of stem and leaves which were dissolved by one-lysis respec-tively were 3.62μg· μL-1 , 4.17μg· μL-1 , and which were dissolved by twi-lysis respectively were 1.53μg·μL-1 ,2.12μg· μL-1 .However ,the protein bandings from SDS-PAGE were all fuzzy ,while the impurities of the pro-tein samples from twi-lysis were fewer than that from one-lysis.The reason possibly was the protein preparation and the electrophoresis condition still must be improved .The research would provide the technical foundation for further-molecular biology studies of agarwood-induction mechanism in Aquilaria sinensis ( Lour) Gilg.
出处
《海峡药学》
2014年第12期37-39,共3页
Strait Pharmaceutical Journal
基金
广东省中药局项目(编号:20122079)
所级项目(编号:2011YZ002)