CAJ | 학술논문

植物液泡膜氢离子焦磷酸酶（Vacuolar H+-PPase,VP）能够酸化液泡并为液泡的次级转运系统提供能量，在植物耐盐性方面发挥着着重要作用。本研究采用RT-PCR结合RACE方法，从费尔干猪毛菜中（Salsola ferganica）克隆得到液泡膜质子焦磷酸酶基因SfVP，该基因的cDNA全长2738 bp，包含1个2301 bp的完整开放读码框（ORF），编码766个氨基酸的多肽。氨基酸序列分析表明其属于H+-转运无机焦磷酸酶超家族成员，具有典型的H+-PPase结构域，其疏水性较强，含有12个跨膜螺旋结构。 SfVP序列比对显示具有H+-PPase的3个保守序列CS1、CS2和CS3，其中CS1中含有保守的底物催化位点，与烟草的NtVP氨基酸序列相似性为95.3％。半定量RT-PCR结果表明，SfVP的转录表达丰度随着600 mmol/L NaCl处理时间的增加而升高，处理12 h后显著增加并一直维持在较高水平。
식물액포막경리자초린산매（Vacuolar H+-PPase,VP）능구산화액포병위액포적차급전운계통제공능량，재식물내염성방면발휘착착중요작용。본연구채용RT-PCR결합RACE방법，종비이간저모채중（Salsola ferganica）극륭득도액포막질자초린산매기인SfVP，해기인적cDNA전장2738 bp，포함1개2301 bp적완정개방독마광（ORF），편마766개안기산적다태。안기산서렬분석표명기속우H+-전운무궤초린산매초가족성원，구유전형적H+-PPase결구역，기소수성교강，함유12개과막라선결구。 SfVP서렬비대현시구유H+-PPase적3개보수서렬CS1、CS2화CS3，기중CS1중함유보수적저물최화위점，여연초적NtVP안기산서렬상사성위95.3％。반정량RT-PCR결과표명，SfVP적전록표체봉도수착600 mmol/L NaCl처리시간적증가이승고，처리12 h후현저증가병일직유지재교고수평。
Plant vacuolar H+-translocating inorganic pyrophosphatase (V-H+-PPase)can acidify vacuoles and power the vacuolar secondary active transport system,which plays an important role in plant salt tolerance.In this study,a vacuolar H+-PPase gene was cloned from Salsola ferganica by RT-PCR and RACE with a 2738 bp full-length cDNA sequence and a 2301 bp open reading frame (ORF)which encoding a putative polypeptide of 766 amino acids.Sequence analysis showed that SfVP protein belonged to the H+-translocating inorganic pyrophosphatase superfamily and had classic H +-PPase multidomain.SfVP protein contains 12 transmembrane structures with strong hydrophobicity.Sequence alignment analysis based on amino acid sequences indicated that SfVP had 3 conservative sequence regions including CS1,CS2 and CS3.CS1 contains conservative substrate binding sites,which showed 95.3% similarity with tobacco NtVP.Semi-quantitative RT-PCR results revealed that transcription level of SfVP increases gradually with the increase of treatment time under 600 mmol/L NaCl stress,and maintains a high express abundance after 12 hour treatment.