芹菜非特异性脂转移蛋白基因的克隆与表达分析  

Cloning and Expression Pattern Analysis of nsLTP Gene in Celery

作　　者：蒋倩[1] 王枫[1] 侯喜林[1] 王镇[1] 李梦瑶[1] 马静[1] 刘梦叠[1] 熊爱生[1] 

机构地区：[1]南京农业大学园艺学院,作物遗传与种质创新国家重点实验室,农业部华东地区园艺作物生物学与种质创制重点实验室,南京210095

出　　处：《园艺学报》2012年第7期1293-1302,共10页Acta Horticulturae Sinica

基　　金：教育部新世纪优秀人才支持计划项目(NECT-11-0670);江苏高校优势学科建设项目(2011PAPD);江苏省双创计划项目(2011JSSC);南京农业大学高层次人才引进项目

摘　　要：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。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： Hla and Hlb, 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.

关 键 词：芹菜 脂转移蛋白 基因克隆 实时定量PCR 基因表达 

分 类 号：S636.3[农业科学—蔬菜学]