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Position: Home > Articles > Purification and Identification of Glutamine Synthetase in Wheat Seedlings Journal of Triticeae Crops 2010,30 (1) 89-92

小麦叶片谷氨酰胺合成酶的分离纯化及鉴定

作  者:
王小纯;安帅;熊淑萍;程振云;陈新建;孟凡荣;马新明
单  位:
河南农业大学农学院;河南农业大学生命科学院
关键词:
小麦;幼苗叶片;谷氨酰胺合成酶;纯化和鉴定
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。
译  名:
Purification and Identification of Glutamine Synthetase in Wheat Seedlings
作  者:
WANG Xiao-chun1,AN Shuai1,XIONG Shu-ping2,CHENG Zhen-yun1,CHEN Xin-jian1,MENG Fan-rong1,MA Xin-ming2 (1.College of Life Science,Henan Agricultural University,Zhengzhou,Henan 45002,China;2.College of Agronomy,Henan Agricultural University,Zhengzhou,Henan 45002,China)
关键词:
Wheat;Seedling leaf;Glutamine synthetase;Gurification and identification
摘  要:
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.

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