【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cDNA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。更多还原

【Abstract】 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 Oglc13Ⅱ 3′end sequence of oat.更多还原