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ID:22399810
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页数:16页
时间:2018-10-29
《丹参蛋白激酶smsnrk2.4的克隆及表达分析》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、丹参蛋白激酶SmSnRK2.4的克隆及表达分析 [摘要]蔗糖非酵解型蛋白激酶家族2(SnRK2)在逆境植物信号转导途径中起着重要作用。该研究根据丹参转录组数据从丹参cDNA中克隆得到一个SnRK2家族基因,命名为SmSnRK24,属于Ⅰ类SnRK2。SmSnRK24基因包含8个内含子,9个外显子,其开放阅读框1068bp,编码355个氨基酸,推测其蛋白相对分子质量为4063kDa,将其构建到原核表达载体pMALc2X上,原核诱导表达出与预测蛋白大小一致的目的蛋白。依据丹参基因组序列,PlantCARE分析SmSnRK2
2、4起始密码子ATG上游3000bp的启动子系列,结果显示该序列包含Ibox,GAmotif,HSE,LTR,TCrichrepeats等逆境胁迫响应元件,以及GAREmotif,Pbox,ABRE,CGTCAmotif等激素响应元件。实时荧光定量PCR分析表明SmSnRK24在丹参根中的含量较高,在茎中和叶中含量相当,ABA和PEG胁迫响应分析表明,SmSnRK24对PEG渗透胁迫响应显著,对ABA胁迫响应微弱。该研究为进一步探讨SmSnRK24基因在丹参干旱胁迫下次生代谢产物积累机制中的作用奠定了基础。 [关键词]丹
3、参;SmSnRK24;原核表达;启动子;实时荧光定量PCR Cloningandexpressionanalysisofproteinkinase SmSnRK24fromSalviamiltiorrhiza JIAYanyan1,RUMei2,JINWeibo3,LIANGZongsuo1,3* (1CollegeofLifeScience,NorthwestAgricultureandForestryUniversity,Yangling712100,China; 2InstituteofSoilandWa
4、terConservation,ChineseAcademyofSciencesandtheMinistryofWaterResources,Yangling712100,China; 3CollegeofLifeScience,ZhejiangSciTechUniversity,Hangzhou310018,China) [Abstract]Sucrosenonfermenting1relatedproteinkinase2(SnRK2)playsakeyroleinabioticstresssignalingin
5、plantsInthisstudy,weclonedaSmSnRK24genebelongingtosubclassIofSnRK2fromSalviamiltiorrhizabyscreeningitstranscriptomedatabaseTheSmSnRK24genecontains8intronsand9exons,witha1068bpopenreadingframeencodingapolypeptideof355aminoacids,thepredictedmolecularmassofwhichis40
6、63kDaProkaryoticexpressionofSmSnRK24proteinusingpMALc2XastheexpressionvectordisplayedthattherecombinantproteinofSmSnRK24geneinEcoliwasconsistentwiththepredictedsizeA3000bppromotersequenceofSmSnRK24containedsomestressresponsiveelementsandhormoneresponsiveelementsQ
7、uantitativerealtimePCRanalysisrevealedthattheexpressionofSmSnRK24inrootwasmuchhigherthanthatinstemandleaf,SmSnRK24wasstronglyinducedbyPEGstress,weaklyinducedbyABAstressThisresearchprovidedabasisforfurtherstudyoftheSmSnRK24geneplayingtheroleinaccumulatemechanismof
8、secondarymetabolitesinSmiltiorrhizaunderdrought [Keywords]Salviamiltiorrhiza;SmSnRK24;prokaryoticexpression;promoter;quantitativerealtimePCR 丹??,为唇形科鼠尾草属多年生草
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