西伯利亚蓼非特异性脂质转移蛋白编码序列的克隆及其盐胁迫下的表达

Cloning of Coding Sequence of Non-specific Transfer Protein from Polygonum sibiricum Laxm.and Expression under Salinity Stress

根据西伯利亚蓼地下茎抑制消减文库(SSH)中获得的非特异性脂质转移蛋白(non-specific lipid transfer protein,nsLTP)EST序列,应用RACE技术克隆了具有PolyA的全长cDNA序列.该序列全长604bp,其5′非翻译区65bp,3′非翻译区227bp,开放阅读框编码103个氨基酸残基;序列分析表明,该基因具有N端信号肽,具有nsLTP家族共有的典型保守区域,属nsLTP家族基因,命名为PsnsLTPs;荧光定量PCR分析表明,PsnsLTPs在西伯利亚蓼叶、茎、地下茎中均有表达.在3%NaHCO3诱导表达下,该基因在地下茎中表达明显受盐胁迫的诱导,推测该基因在抵御盐胁迫时具有重要作用.

The full-length nsLTP, a non-specific lipid transfer protein gene, was cloned using RACE （rapid amplification of cDNA end） technology based on a nsLTP partial sequence obtained from a random clone in SSH library of Polygonum sibiricum Laxm. The acquired 604 bp sequence includes a 5＇ untranslated region of 65 bp, a 3＇ untranslated region of 227 bp with poly （A）, and an open reading frame （ORF） encoding 140 amino acids. The sequencing result revealed that the gene, we termed as PsnsLTPs, contained a N-terminal signal peptide that was a typical conservative region of the nsLTP family. The expression analyses by RT-PCR showed that PsnsLTPs was expressed in leaves, stems and rhizomes of Polygonum sibiricum Laxm. Under the induction of 3 % NaHCO3, the expression of PsnsLTPs was significantly increased, which suggested that PsnsLTPs might play an important role in saline stress resistance.