小麦谷氨酰胺合成酶基因克隆与其表达特性分析

摘要

In order to study the function of glutamine synthetase ( GS) in wheat, the full-length cDNA of GS1 and GS2 were cloned by RACE and RT-PCR (The accession number of GSl is HQ840647, and GS2 is JF894116). The full length cDNA of GSl is 1 494 bp and that of GSl is 1 631 bp. The sequences and protein characteristics of GSl and GS2 were analyzed with the biological software. Semi-quantitative PCR and western-blot were used to demonstrate the transcriptional and expressional levels of GS genes in wheat leaf during seedling stage, and the results showed that both levels of GSl got higher when the leaf began to senescence while both levels of GS2 kept the highest from the leaf expanding stage to the full expansion stage.%为了研究小麦谷氨酰胺合成酶(Glutamine synthetase,GS)的功能,利用RACE及RT-PCR技术克隆了小麦GS1(GenBank登录号:HQ840647)和GS2(GenBank登录号:JF894116)基因的全长cDNA.小麦GS1的cDNA全长为1 494 bp,GS2的cDNA全长为1 631 bp,并利用生物信息软件对GS1和GS2的基因序列以及所表达的蛋白特性进行了分析.利用半定量PCR和Western-bolt技术分别分析了小麦苗期叶片发育过程中GS同工酶基因转录和表达特点,结果表明,GS1在叶片开始衰老时转录和表达水平较高,GS2从叶片伸长期到定长期转录和表达水平最高.