沙柳SpsLAS基因克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of SpsLAS Gene in Salix psammophila

作　　者：路东晔 贺玉娇 金娜[1] 刘乐[1] 卜松雪 王雷[1] 杨海峰[1] Lu Dongye He Yujiao Jin Na Liu Le Bu Songxue Wang Lei Yang Haifeng(Forestry College of Inner Mongolia Agricultural University, Hohhot, 010019)

机构地区：[1]内蒙古农业大学林学院,呼和浩特010019

出　　处：《分子植物育种》2017年第2期483-491,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31360187);林木遗传育种国家重点实验室(中国林业科学研究院)开放课题基金(TGB2015001)共同资助

摘　　要：LAS基因对于植物侧生分生组织有重要影响。它被证实在拟南芥、番茄等草本科植物营养生长阶段参与腋生分生组织的形成,但在木本植物中的基因功能尚缺乏深入研究。本研究以沙柳为材料,利用RT-PCR技术成功克隆沙柳LAS基因,命名为SpsLAS,并对其进行生物信息学分析。结果表明,该基因CDS序列全长1 320 bp,编码439个氨基酸,预测蛋白分子量为49.876 8 k D,理论等电点(PI)为6.66,是亲水性蛋白;同源氨基酸序列比对结果表明沙柳LAS基因与番茄LS、拟南芥LAS等功能已确认的LS亚家族基因同源性较高,氨基酸相似性达50%以上;系统进化分析表明SpsLAS与杞柳和毛果杨亲缘关系更近;对LAS进行功能结构域分析得知LAS基因具有GRAS基因家族典型特征,属于GRAS基因家族LS亚家族;预测未发现跨膜结构和信号肽区域;SpsLAS蛋白二级结构包含173个α螺旋(Alpha helix),72个延伸链(Extended strand),194个无规则卷曲结构(Random coil);亚细胞定位预测该基因很可能定位于细胞核内。沙柳LAS基因的克隆对于研究木本植物萌蘖及其分枝机制有重要意义,同时丰富了木本植物中GRAS基因家族的相关研究。The LAS genes play very important roles in plant lateral meristem development. It was confirmed that those genes take part in axillary meristem development during vegetative phase in Arabidopsis, tomato and other herb plants, but the function of these genes in woody plants is still in a lack of further study. In this paper, Salix psammophila was used as materials, LAS gene was cloned successfully by RT-PCR method and named SpsLAS.Bioinformatics analysis were carried out. The results showed that the full-length CDS of LAS was 1 320 bp, and encoded the protein which consists of 439 amino acids. The putative protein molecular weight was 49.876 8 Da and its theoretical isoelectric points was 6.66, SpsLAS was a hydrophilic protein; Homologous amino acid sequence comparison showed that the SpsLAS gene had higher homology with LS subfamily genes which gene function had been identified, like LS gene in tomato, LAS gene in Arabidopsis,the similarities of putative amino acid sequences of the cloned LAS genes were over 50%; Phylogenetic analysis showed that Salix psammophila,Salix purpurea and Populus trichocarpa were closely related; Functional domain analysis showed that LAS gene has the typical characteristics of GRAS gene family and belongs to the LS subfamily; There was no transmembrane domain and signal peptide region in SpsLAS; The secondary structure of protein contained 173 alpha helixs, 72 extended strands, 194 random coils; The subcellular localization prediction of the gene was likely to locate in the nucleus. Cloning of SpsLAS gene has important significance for the research of woody plants on shoot branching mechanism, but also enrich the correlation of GRAS gene family members in woody plants.

关 键 词：沙柳 LAS 基因克隆 生物信息学分析 

分 类 号：S793.9[农业科学—林木遗传育种] Q943.2[农业科学—林学]