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以芹菜(Apium graveolens)‘六合黄心芹’、‘津南实芹’和‘美国西芹’为试验材料,采用RT-PCR技术分别获得其cDNA序列。序列分析表明:来源于3个芹菜品种的非特异性脂转移蛋白(Non-specific lipid transfer protein,nsLTP)基因核苷酸序列高度保守,全长357bp,编码118个氨基酸,起始密码子ATG之后含有27个氨基酸残基的信号肽序列,推测其成熟的蛋白含91个氨基酸残基,预测其蛋白质分子量为11.75kD,pI值为9.36。芹菜的nsLTP蛋白主要由α-螺旋和随机卷曲组成。空间结构上分析显示,芹菜nsLTP蛋白中H1区域明显分为H1a和H1b两个亚区域,而模板碧桃中H1区域为一个连续的螺旋结构,存在明显的差异。进化分析显示,芹菜nsLTP与香石竹、大洋洲滨藜等植物的nsLTP相似性较高,在保守位置具有8个半胱氨酸残基。实时定量PCR表达分析表明,该基因主要在芹菜的茎以及茎尖生长活跃中心表达,具有明显的组织特异性。
The cDNA sequences were obtained by RT-PCR using Apium graveolens ’Liuhe Huangxinqin’, ’Jinnan Shiqin’ and ’American Celery’ as test materials. Sequence analysis showed that the nucleotide sequence of the non-specific lipid transfer protein (nsLTP) gene derived from three celery cultivars was highly conserved, with a total length of 357 bp, encoding a polypeptide of 118 amino acids. After initiation codon ATG, 27 amino acid residue signal peptide sequence, suggesting that the mature protein contains 91 amino acid residues, the predicted molecular weight of 11.75kD, pI value of 9.36. The celery’s nsLTP protein consists mainly of alpha-helices and random curls. The spatial structure analysis showed that the H1 region of celery nsLTP protein was divided into two sub-regions of H1a and H1b, while the H1 region of the template was a continuous helical structure with obvious differences. Phylogenetic analysis showed that the nsLTP of celery nsLTP had similarities with nsLTP of plants such as Carnation and Oceania, and had 8 cysteine residues in the conserved position. Quantitative real-time PCR analysis showed that the gene was mainly expressed in the stems and tips of celery shoots, and had obvious tissue specificity.