YU Wenjing,LIU Zhihua,DIAO Guiping,et al.Cloning and Prokaryotic Expression Vector Construction of Eliciting Plant Response Protein Gene TatEpl1[J].Northern Horticulture,2015,39(16):93-97.[doi:10.11937/bfyy.201516022]
刺激植物响应蛋白基因TatEpl1的克隆及原核表达载体构建
- Title:
- Cloning and Prokaryotic Expression Vector Construction of Eliciting Plant Response Protein Gene TatEpl1
- 关键词:
- 深绿木霉ACCC30153; 刺激植物响应蛋白; 原核表达
- 文献标志码:
- A
- 摘要:
- 以深绿木霉（Trichoderma atroviride）ACCC30153为试材，采用PCR技术克隆到刺激植物响应蛋白基因TatEpl1，并构建TatEpl1的原核表达载体。结果表明：经测序得到的cDNA和DNA序列长度分别为417 bp和487 bp，接受号分别为JN695780和JN695781。以cDNA为模板进行PCR获得TatEpl1基因片段，并将目的片段插入原核表达载体pGEX-4T-2的相应位置，获得重组表达载体pGEX-TatEpl1，并将其转入大肠杆菌（Escherichia coli）BL21中获得重组菌株BL21-TatEpl1，经检测均呈阳性。
- Abstract:
- Using the T.atroviride ACCC30153 as material，the eliciting plant response protein gene TatEpl1 was cloned by PCR，and the prokaryotic expression vector of TatEpl1 was constructed.The results showed that,the cDNA sequence of TatEpl1 was 417 bp in length，and the DNA sequence was 487 bp in length and contained two exons and one intron.The cDNA and DNA sequences of TatEpl1 were deposited in the GenBank database with the accession numbers JN695781 and JN695780，respectively.The segment of TatEpl1 was inserted into prokaryotic expression vector pGEX-4T-2 and transformed into E.coli BL21 for obtaining the recombinant strain BL21-TatEpl1.The results showed all of the transformants were positive by detection.
参考文献/References:
[1]王芳.深绿木霉的拮抗机理、促生作用及制剂加工研究[D].兰州：甘肃农业大学，2006. [2]徐秉良.不同环境对深绿木霉菌株T2抑制黄瓜黑星病菌的影响[C].中国植物病理学会2004年学术年会论文集，2004：453-456. [3]DUBEY M K，BROBERG A，SOORIYAARACHCHI S，et al.The glyoxylate cycle is involved in pleotropic phenotypes，antagonism and induction of plant defence responses in the fungal biocontrol agent Trichoderma atroviride[J].Fungal Genet Boil，2013，58-59:33-41. [4]RUOCCO M，LANZUISE S，VINALE F，et al.Idntification of a new biocontrol gene in Trichoderma atroviride：the role of an ABC transporter membrane pump in the interaction with different plant-pathogenic fungi [J].Mol Plant Microbe Interact，2009，22:291-301. [5]孙健，王志英，刘志华，等.深绿木霉发酵液对山新杨防御酶活性的影响[J].安徽农业科学，2014，42(32):11344-11345. [6]MING Q，SU C，ZHENG C，et al.Elicitors from the endophytic fungus Trichoderma atroviride promote Salvia miltiorriza hairy root growth and tanshinone biosynthsis [J].J Exp Bot，2013，64:5687-94. [7]DJONOVI〖KG-*3〗C〖DD(-*2/3〗′〖DD)〗S，POZO M J，DANGOTT L J，et al.Sm1，a Proteinaceous elicitor secreted by the biocontrol fungus Trichoderma virens induces plant defense responses and systemic resistance[J].Molecular Plant-Microbe Interactions,2006,19(8):838-853. [8]DJONOVIC S，VARGAS W A，KOLOMIETS M V，et al.A proteinaceous elicitor sm1 from the beneficial fungus trichoderma virens is required for induced systemic resistance in maize[J].Plant Physiol,2007,145(3):875-889. [9]王允.棘孢木霉和球毛壳菌诱导蛋白及其功能研究[D].哈尔滨：哈尔滨工业大学,2013. [10]SEIDL V，MARCHETTI M，SCHANDL R，et al.Epl1，the major secreted protein of Hypocrea atroviridis on glucose，is a member of a strongly conserved protein family comprising plant defense response elicitors[J].FEBS J，2006,73:4346-4359. [11]古丽吉米拉·米吉提.棘孢木霉刺激植物响应蛋白基因Epl1功能研究[D].哈尔滨：东北林业大学，2013. [12]王全永，黄世文，王玲，等.我国植物激活蛋白诱导的生物化学活研究进展及应用[J].植物保护，2007(4):20-22.
备注/Memo
第一作者简介：遇文婧（1984-），女，黑龙江哈尔滨人，博士研究生，现主要从事森林保护等研究工作。E-mail：ywjlinda2008@163.com. 责任作者：王志英（1956-），男，黑龙江哈尔滨人，硕士，教授，现主要从事森林保护等研究工作。E-mail：WZYNEFU@126.com. 基金项目：国家自然科学基金面上资助项目（NSFC：31170601）；黑龙江省青年科学基金资助项目（QC2011C003）；黑龙江省教育厅科学技术研究资助项目（12513024）；黑龙江省级基本科研业务费资助项目（省财政自拟项目）。