粗糙脉孢菌snRNA基因的克隆及其表达调控研究  

The Cloning and characterization of Neurospora crassa sn RNA genes

作　　者：万宇峰[1] 郑桂娜[1] 张琳[1] 沈寒婕 陈现云 王东妮[1] 郭金虎[1] 

机构地区：[1]中山大学生命科学学院有害生物控制与资源利用国家重点实验室,广东广州510006

出　　处：《菌物学报》2015年第1期98-107,共10页Mycosystema

基　　金：国家973课题(2011CB711000和2012CB47600);国家自然科学基金面上项目(31071122和31171119);高等学校博士学科点专项科研基金(20110171110030);教育部新世纪优秀人才支持计划(NCET-12-0566)

摘　　要：Pre‐m RNA(precursor m RNA)的剪接是真核基因表达中的重要一环,由剪接体复合物(spliceosome)催化完成。小核RNA(small nuclear RNAs,sn RNAs)是剪接体的重要结构和功能组分。本工作首次鉴定了粗糙脉孢菌的U1、U2、U4、U5和U6等sn RNA基因,这些基因除U5为单一拷贝外,其余为多拷贝基因且表达量存在差异。对各基因的近端序列元件(proximal sequence elements,PSEs)的分析显示在大部分基因都存在一段回文的保守序列GTGCAC,荧光素酶报告基因实验证实该序列具有调控部分sn RNA基因转录的功能。我们还通过温度梯度实验检测了stk‐16第三内含子的剪接情况变化,结果提示可变剪接对调节生物可能对不同温度环境的适应具有重要作用。Pre‐mRNA splicing plays an important role in regulating eukaryotic gene expression, which is mediated by the spliceosome complex. sn RNAs are crucial components for the structure and function of spliceosome. In this work, we identified the sn RNA genes from Neurospora crassa, which included U1, U2, U4, U5 and U6, and each comprised various variant numbers with the exception of U5. These sn RNA gene variants show different expression levels. Analysis of the proximal sequence elements indicated that a palindromic consensus sequence GTGCAC existed in most of these sn RNA genes. The regulation of some sn RNA genes transcribed by this sequence was confirmed via luciferase assay. We further examined the stk‐16＇s intron‐3 alternative splicing in temperature gradient, and the results suggest that alternative splicing may have a role in climate temperature change adaptation.

关 键 词：粗糙脉孢菌 小核RNA 可变剪接 近端序列元件 stk‐16基因 

分 类 号：Q78[生物学—分子生物学]