关键词: 芹菜, 脂转移蛋白, 基因克隆, 实时定量PCR, 基因表达

Abstract: In this study，full-length of cDNA sequences of non-specific lipid transfer protein （nsLTP）gene were cloned from celery（Apium graveolens）cultivars‘Liuhe Huangxinqin’，‘Jinnan Shiqin’and ‘Meiguo Xiqin’using reverse transcript PCR（RT-PCR）. Sequence analysis shows：The cDNA nucleotide sequences are highly conserved from the three cultivars. The length of the gene is 357 bp，containing a complete open reading frame to encode 118 amino acids. There is a signal peptide sequence with 27 amino acid residues. The mature protein contains 91 amino acid residues. Its molecular mass is 11.75 kD，and pI is 9.36. Amino acid sequence comparison indicates that the nsLTP from celery has a high similarity with the nsLTPs from Dianthus caryophyllus and Atriplex nummularia. There are 8 Cys amino acid residues in the conservative position. The nsLTP protein from celery is mainly composed by α-helixs and random coils. Spatial structure analysis shows significant differences. The H1 region of nsLTP protein from celery was divided into two sub-region：H1a and H1b，while the H1 region of template is a continuous helical structure. Quantitative real-time PCR analysis shows that the gene is tissue-specific and mainly expressed in the stem and active center of shoot apex in celery.

Key words: Apium graveolens, nsLTP, gene clone, quantitative real-time PCR, gene expression