小麦叶片谷氨酰胺合成酶的分离纯化及鉴定  

Purification and Identification of Glutamine Synthetase in Wheat Seedlings

作　　者：王小纯[1] 安帅[1] 熊淑萍[2] 程振云[1] 陈新建[1] 孟凡荣[1] 马新明[2] 

机构地区：[1]河南农业大学生命科学院,河南郑州450002 [2]河南农业大学农学院,河南郑州450002

出　　处：《麦类作物学报》2010年第1期83-86,共4页Journal of Triticeae Crops

基　　金：国家自然基金项目(30771266);农业部公益项目(2007-05)

摘　　要：为了解谷氨酰胺合成酶(Glutamine synthetase,GS)同工酶在小麦中的种类、活性及亚基组成状况,利用盐析、凝胶过滤、阴离子交换层析等技术对小麦苗期叶片中的GS同工酶进行了分离纯化及鉴定。通过测定GS活性跟踪GS的纯化过程,利用Native-PAGE结合活性染色技术检测GS的纯化效果,利用SDS-PAGE鉴定GS的亚基组成。结果表明,在小麦幼苗叶片中检测到3种GS同工酶GS1、GS2和GSx,其中Gsx含量少,但活性高。从小麦幼苗叶片中纯化得到2个GS同工酶即GS1和GS2。GS1由39 kD的亚基组成,GS2由45 kD的亚基组成,没有纯化得到GSx。To confirm the role of glutamine synthetase isoforms in the nitrogen assimilation of wheat, three glutamine synthetase isoforms, GS1, GS2 and GSx in the wheat leaf during seedling period were purified with salting-out, gel filtration chromatography and anion-exchange chromatography. The purification process was traced by determining the GS activity of each fraction, and the purification effect was examined by Native-PAGE combined with activity staining. The subunit component of GS isoforms was analyzed by SDS-PAGE. The result showed that GS1 and GS2 was purified from leaves of wheat seedlings. GS1 was composed with a 39 kD subunit, and GS2 was composed with a 45 kD subunit. GSx was not obtained by this purification process.

关 键 词：小麦 幼苗叶片 谷氨酰胺合成酶 纯化和鉴定 

分 类 号：S512.1[农业科学—作物学] S312