燕麦β-1,3-葡聚糖酶Ⅱ基因3′端cDNA的克隆及分析  

Cloning and Sequence Analysis of β-1,3-glucanase Ⅱ Gene 3′End cDNA of Oat

作　　者：何江峰[1] 韩冰[1] 郭慧琴[1] 张占雄[1] 赵雅丽[1] 

机构地区：[1]内蒙古农业大学生物工程学院,内蒙古呼和浩特010018

出　　处：《生物技术》2007年第1期5-8,共4页Biotechnology

基　　金：高校基金项目资助("燕麦胚乳发育β-1;3葡聚糖的积累与调控";编号:K32608)

摘　　要：实验利用3＇-RACE技术进行燕麦β-1,3-葡聚糖酶Ⅱ（Oglc13Ⅱ）3＇末端cDNA的快速扩增并进行序列分析.将0.01%的HgCl2诱导处理24h之后的燕麦（Avena Sativa）幼叶作为材料,利用RT-PCR技术得到Oglc13Ⅱ的部分cDNA片段,根据此片 段设计一条特异性上游引物,反转录引物中的部分序列即3＇sites Adaptor Primer作为下游引物,按3＇RACE试剂盒（Takara）操作流程进行Oglc13Ⅱ3＇末端cDN A的快速扩增,成功获取837bp的3＇-RACE反应产物,通过BLAST技术,发现该片段与 许多植物β-1,3-葡聚糖酶基因具有较高的相似性（50.0%～72.0%）.因此认为成功 克隆了Oglc13ⅡcDNA的3＇末端序列.To amplify the 3′-cDNA end of oat β-1,3-glucanase Ⅱ（oglc13Ⅱ） by 3′-RACE technique and to analyze the sequence in the experiment.A part of oat Oglc13Ⅱ partial cDNA fragment was amplified by RT-PCR from total RNA of oat′s leaves which had been induced for 24 hours by HgCl2（0.01%）firstly.We designed a special primer on this fragment.With this special primer as up primer and 3′sites Adaptor Primer as down primer,3′-cDNA end of oglc13Ⅱ was rapid amplified by following the instruction of 3′RACE kit（Takara）. 3＇ - RACE product with 837bp was successfully obtained, Homology analysis by BLAST showed highly homologous（50.0% - 72.0） with β- 1,3- glucanase of other plants. So the sequence was thought as the Oglc13 Ⅱ 3＇end sequence of oat.

关 键 词：β-1 3-葡聚糖酶Ⅱ基因 RT-PCR 3'RACE CDNA 

分 类 号：Q523[生物学—生物化学] Q785