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当前位置: 首页 > 文章 > 小麦TaSPL17基因的克隆、组织特异性表达及原核表达分析 西北农业学报 2015,24 (7) 37-43
Position: Home > Articles > Cloning,Tissue-specific Expression and Prokaryotic Expression Analysis of TaSPL17 Gene in Wheat(Triticum aestivum) Acta Agriculturae Boreali-occidentalis Sinica 2015,24 (7) 37-43

小麦TaSPL17基因的克隆、组织特异性表达及原核表达分析

作  者：
张铁怀;徐开杰;刘曙东;奚亚军;孙风丽
单  位：
西北农林科技大学农学院
关键词：
小麦(Triticum aestivum);TaSPL17;基因克隆;组织特异性表达;原核表达
摘  要：
具有SBP结构域的SPL(SQUAMOSA promoter-binding protein-like)蛋白是一类植物特有的转录因子,在植物的生长发育过程中发挥重要的调控作用。为了进一步研究小麦基因TaSPL17的功能,本研究通过同源克隆的方法,从普通小麦品种‘小偃22’中克隆得到TaSPL17基因全长CDS序列。生物信息学分析表明,该基因开放阅读框长度1 158bp,编码385个氨基酸,理论分子质量40.25ku,理论等电点为9.12。半定量RT-PCR结果显示,TaSPL17在小麦根、茎基部、叶片、幼穗中均有表达,在幼穗和茎基部中表达量最高,根次之,在叶片中表达量最少,表明TaSPL17基因可能与小麦的分蘖和穗部的发育有关。将基因TaSPL17与载体pGEX6P-1连接构建原核重组表达载体pGEX6P-1-TaSPL17,然后将其转入大肠杆菌DH5α并对诱导表达的时间、IPTG浓度、温度进行优化,SDS-PAGE分析表明,融合GST标签的TaSPL17在温度37℃,IPTG浓度为0.4mmol/L,诱导3h后表达量最大,有助于进一步纯化TaSPL17蛋白质。
译  名：
Cloning,Tissue-specific Expression and Prokaryotic Expression Analysis of TaSPL17 Gene in Wheat(Triticum aestivum)
作  者：
ZHANG Tiehuai;XU Kaijie;LIU Shudong;XI Yajun;SUN Fengli;College of Agronomy,Northwest A&F University;Key Laboratory of Wheat Biology and Genetic Improvement on Northwestern China,Ministry of Agriculture;
关键词：
Wheat;;TaSPL17;;Gene cloning;;Tissue-specific expression;;Prokaryotic expression
摘  要：
SPL(SQUAMOSA promoter-binding protein-like)proteins with SBP box character is a family of plant-specific transcription factors,which plays important roles in the process of plant growth and development.To further investigate the function of the TaSPL17 gene,we obtained a-1182 bp fragment includes the TaSPL17 gene from common wheat variety Xiaoyan 22 by using method of homologous cloning.Bioinformatics analysis showed that the gene TaSPL17 with a 1 158 bp length open reading frame,encoded a protein contained 385 amino acids.The predicted pI of TaSPL17 was9.12 and molecular mass 40.25 ku.The results of semi-quantitative RT-PCR showed that the gene in stem base and young ear were higher than in roots and leaves.These results indicated that the TaSPL17 gene may play important roles in the development of tillers and panicles.The gene TaSPL17 connected with;expression vector pGEX6P-1to construct prokaryotic recombinant expression vectorpGEX6P-1-TaSPL17.This recombinant expression vector was transformed into E.coli DH5α,then optimized the time,IPTG concentration and temperature of recombinant plasmid induction expression.Analysis of SDS-PAGE showed that concentration of IPTG was 0.4mmol/L at 37 ℃ of fusion protein GST,the optimal condition for the expression was induced for 3h,this will help to further purification TaSPL17 protein.This work lays a foundation for the further research of TaSPLgenes regulation in wheat plant architecture development,and will help to reveal the molecular mechanism of TaSPL17 regulating wheat architecture and yield traits.

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