CAJ | 학술논문

从普通烟草腺毛cDNA文库中筛查烟草SnRK2基因的EST序列，以此序列为信息探针，通过电子克隆和RT-PCR的方法从烟草中克隆到一个包含1017 bp开放阅读框、编码338个氨基酸的cDNA序列，命名为NtSnRK2.1。生物信息学分析表明， NtSnRK2.1蛋白同时具有磷酸化丝氨酸/苏氨酸和酪氨酸的活性。氨基酸序列比对发现，NtSnRK2.1与拟南芥、水稻和小麦等植物中受逆境胁迫诱导表达的直系同源基因高度同源。组织表达分析结果显示，烟草NtSnRK2.1基因的组织表达差异较大，在根部的表达量最高，其次是叶片，茎部的表达量最低。胁迫处理下的基因表达分析结果表明，NtSnRK2.1受高盐、高渗、低温胁迫和ABA处理诱导表达，对各胁迫条件的应答模式不同，其对各胁迫条件的敏感度为：高渗＞高盐＞低温＞ABA。
종보통연초선모cDNA문고중사사연초SnRK2기인적EST서렬，이차서렬위신식탐침，통과전자극륭화RT-PCR적방법종연초중극륭도일개포함1017 bp개방열독광、편마338개안기산적cDNA서렬，명명위NtSnRK2.1。생물신식학분석표명， NtSnRK2.1단백동시구유린산화사안산/소안산화락안산적활성。안기산서렬비대발현，NtSnRK2.1여의남개、수도화소맥등식물중수역경협박유도표체적직계동원기인고도동원。조직표체분석결과현시，연초NtSnRK2.1기인적조직표체차이교대，재근부적표체량최고，기차시협편，경부적표체량최저。협박처리하적기인표체분석결과표명，NtSnRK2.1수고염、고삼、저온협박화ABA처리유도표체，대각협박조건적응답모식불동，기대각협박조건적민감도위：고삼＞고염＞저온＞ABA。
One EST ofSnRK2was screened from glandular trichome cDNA library of tobacco. Based on the EST sequence,NtSnRK2.1 was isolated from tobacco (Nicotiana tabacum L.) byin silicocloning and RT-PCR.NtSnRK2.1 includes an open reading frame (ORF) of 1017 bp and encodes 338 deduced amino acid residues (AAR) with a calculated molecular mass of 43 kDa and a predicted pI of 5.78. Scansite analysis indicated that NtSnRK2.1 contained potential serine/threonine protein kinase activities like other SnRK2 family members. Phylogenetic analysis suggestedNtSnRK2.1 was high homologous to its orthologous genes from Arabidopsis, rice and wheat, which were also induced by abiotic stresses. Reverse transcription-polymerase chain reaction (RT-PCR) and quantitative real-time PCR (qRT-PCR) was used to determine expression patterns ofNtSnRK2.1 in tobacco. Results revealed thatNtSnRK2.1 expressed most strongly in tobacco roots, more in leaves, and marginally in stems. Expression patterns under abiotic stress responses suggested thatNtSnRK2.1 was involved in response to NaCl, PEG, cold stresses and ABA treatment, with significant different responsive profiles. The sensitivity degrees of NtSnRK2.1 responding to four treatments was in the order of hyperosmolality> high salinity> low temperature (4℃)> abscisic acid.