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通过BLAST比对分析丹参EST数据库,发现一个与普遍胁迫蛋白(USP)基因家族同源性较高的基因序列,命名为SmUSP,GenBank登录号为JX416284。依据该序列,从DNA和cDNA水平克隆得到该基因全长,经分析发现该基因无内含子序列,包含一个长711bp的完整开放读码框(ORF),编码236个氨基酸残基。生物信息学分析显示,SmUSP编码蛋白的相对分子量25.5kDa,为无信号肽和跨膜结构域的疏水酸性蛋白;SmUSP具有UspA结构域和ATP结合位点,属于USP蛋白UspFG亚家族。实时荧光定量PCR分析表明,SmUSP在丹参的根、茎、叶、花中均有表达,在根中的表达量较高。同时,SmUSP表达受茉莉酸甲酯(MeJA)、水杨酸(SA)、脱落酸(ABA)以及脱水胁迫的上调诱导,推测该基因可能在丹参防御反应中发挥作用。
By BLAST analysis of Salvia miltiorrhiza EST database, a gene sequence homologous to the common stress gene (USP) gene family was found and named as SmUSP. GenBank accession number was JX416284. Based on this sequence, the full length of the gene was cloned from the DNA and cDNA. The gene was found to contain no intron sequence and contained a 711bp complete open reading frame (ORF) encoding 236 amino acid residues. Bioinformatics analysis showed that the relative molecular mass of SmUSP-encoded protein was 25.5 kDa, which was a hydrophobic acidic protein with no signal peptide and transmembrane domain. SmUSP had UspA domain and ATP binding site and belonged to the UspFG subfamily of USP protein. Real-time PCR analysis showed that SmUSP was expressed in root, stem, leaf and flower of Salvia miltiorrhiza and higher in roots. At the same time, SmUSP expression was induced by MeJA, SA, ABA and dehydration stress, suggesting that this gene may play a role in the defense response of Salvia miltiorrhiza.