金发草GDP-D-甘露糖焦磷酸化酶基因的克隆及功能分析  

Cloning and analysis of GDP-D-mannose pyrophosphorylase gene from Pogonatherum paniceum

作　　者：范林洪[1] 李锐[1] 童永鳌[1] 王胜华[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2015年第3期682-688,共7页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然基因(3107276;31270360)

摘　　要：通过RACE-PCR技术克隆得到金发草GMP基因的cDNA全长序列,命名为PpGMPase(GenBank序列号:KF586841).该基因开放阅读框长度为1086bp,编码361个氨基酸,分子式为C1787H2874N474O509S17;DNA序列由4个外显子和3个内含子组成;该基因与玉米、水稻、猕猴桃、烟草、拟南芥、番木薯等植物GMP基因具有较高的同源性,与二穗短柄草亲缘关系最近.采用荧光定量PCR方法对该基因响应非生物胁迫(盐、干旱和冷胁迫)的表达模式进行分析,结果表明:在高盐、干旱及低温胁迫后GMP基因的表达量都有显著性增加,并且其催化产物抗坏血酸含量也随之增加.A full-length GDP-D-mannose pyrophosphorylase （GMP） cDNA from Pogonatherurn panice- urn was cloned through RACE method using known GMP gene fragments and named PH-GMPase （GeneBank Accession Number： KF586841）. The PH-GMPase ORF contains 361 amino acids and the molecular formula of it is C1787H2874N4740509S17. It＇s DNA Sequence is consist of 4 exons and 3 in- trons. The sequence homology analysis shows PH-GMPase has a 98.06%, 93.07%, 88.64%, 86.98%, 87.53% and 86.43% similarity with Zea rna ys , Oryza Sativa , Actinidia chinensis , Nicotiana tabacurn , Arabidopsis thaliana and Manihot esculenta, respectively. The constructed phylogenetic tree indicates PH-GMPase has a common ancestor with other higher plants, fungi, higher animals and bacteria and has a nearest relationship with Brachypo diurndistachyon. The expression level of GMP was determined un- der salt, drought and cold stress by fluorescent quantization. The result showed GMP was up-regulated in both three stress conditions, and the content of ascorbic acid （AsA） subsequently increased.

关 键 词：金发草 GDP-D-甘露糖焦磷酸化酶 抗逆性 抗坏血酸 基因克隆 

分 类 号：Q949.7[生物学—植物学]