芹菜中6-磷酸甘露糖还原酶基因的克隆、进化和表达分析  

Cloning,evolution and expression pattern analysis of M6PR gene in celery

作　　者：蒋倩[1] 王枫[1] 侯喜林[1] 马静[1] 李梦瑶[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,江苏南京210095

出　　处：《南京农业大学学报》2013年第4期18-24,共7页Journal of Nanjing Agricultural University

基　　金：教育部新世纪优秀人才支持计划项目(NCET-11-0670);国家自然科学基金项目(31272175);江苏高校优势学科建设项目(2011PAPD);江苏省双创计划项目(2011JSSC)

摘　　要：以2个芹菜品种‘津南实芹’和‘美国西芹’为试材,采用RT-PCR技术分别获得6-磷酸甘露糖还原酶(M6PR)cDNA序列。序列分析表明:来源于2个芹菜品种的6-磷酸甘露糖还原酶基因核苷酸序列全长均为930 bp,编码309个氨基酸;预测其蛋白质相对分子质量为35×103,pI值为6.38。空间结构分析显示:M6PR蛋白由11个α-螺旋和13条β-延伸主链组成,中间形成1个疏水穴;催化四分体(Asp、Tyr、Lys和His)分布于疏水穴内部,具有催化活性。进化分析显示:芹菜M6PR与卷柏、小立碗藓、云杉等远古物种的醛-酮还原酶(AKR)相似性较高。实时定量PCR表达分析表明:M6PR基因主要在芹菜的茎中表达,具有明显的组织特异性。Full-lengths of cDNA sequences encoding mannose-6-phosphate reductase(M6PR)were cloned from celery(Apium graveolens)cultivars'Jinnanshiqin'and'Meiguoxiqin'with reverse transcript PCR(RT-PCR).Sequence analysis showed that the length of the gene was 930 bp,containing a complete open reading frame,which encoded 309 amino acids residues.Its relative molecular mass was 35×103,and pI was 6.38.Three-dimension structure analysis showed that M6PRs from the two celery cultivars were composed by 11 α-helices and 13 β-strands,among which there was a hydrophobic cavity.Catalysis was promoted by a conserved tetrad of active site residues(Tyr,Lys,Asp and His).Amino acid sequence comparison indicated that the M6PRs from'Jinnanshiqin'and'Meiguoxiqin'had high similarity with the AKRs from Selaginella moellendorffii,Physcomitrella patens and Picea sitchensis.Quantitative real-time PCR analysis showed that M6PR gene was tissue-specific and was mainly expressed in the stem.

关 键 词：芹菜 6-磷酸甘露糖还原酶 基因克隆 进化 实时定量PCR 基因表达 

分 类 号：S636.3[农业科学—蔬菜学]