枣果实β-半乳糖苷酶基因的克隆及表达分析  

Cloning and Expressing Analysis of β-galactosidase Gene in Jujube Fruit

作　　者：吕燕荣[1] 任小林[1] 周会玲[1] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨陵712100

出　　处：《西北植物学报》2011年第7期1318-1325,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家苹果产业技术体系专项(nycytx-08-05-02)

摘　　要：采用3-′RACE技术获得了梨枣果实β-半乳糖苷酶(-βGal)基因的部分cDNA片段,并运用实时荧光定量PCR技术和PNPG反应比色法,研究了梨枣不同生长期及采后常温贮藏过程中ZJGAL基因的相对表达量和β-Gal活性的变化,以探索β-半乳糖苷酶在枣(Ziziphus jujubaMill.)果实生长及采后软化中的分子调控机制.结果表明,克隆的-βGal基因序列长2 584 bp(GenBank登录号HQ827769),其中包含2 193 bp的最大阅读框,编码730个氨基酸,将该基因命名为ZJGAL.Blastn和Blastp序列比对分析发现,该ZJGAL基因与已登录的其他物种的β-Gal基因相似性达到60%～80%.随着果实的生长、成熟及采后衰老,ZJGAL基因的表达量和-βGal活性在果实采收前后均呈先上升后下降的趋势,两者的采前高峰均在幼果膨大期,但是采后贮藏过程中虽然基因的表达量较高,但是-βGal活性一直较低.In order to explore the molecular regulation mechanism of β-galactosidase（β-Gal） in ‘Lizao’（Ziziphus jujuba Mill.）fruit,a partial β-Gal cDNA sequence was obtained by 3′-RACE,the expression of the gene and β-Gal activity during fruit growth and ripening at the room temperature were investigated by real-time quantitative PCR（RT-qPCR） approach and PNPG reaction method.The results showed that,the partial β-Gal cDNA sequence named ZJGAL was 2 584 bp（GenBank accession No.HQ827769）,and it contained 2 193 bp ORF and coded 730 amino acid.The results of sequence alignment analysis by Blastn and Blastp in GenBank demonstrated that this β-Gal sequence had 60%～80% identities on the nucleotide sequence and the deduced amino acid sequence to those of other plants.The expression of ZJGAL gene and β-Gal activity displayed the same increasing firstly and then decreasing trend during the fruit growth and ripening,and both of the maximum during the fruit growth was reached at the young fruit expanding period.Even though the expression of ZJGAL gene maintained a high level during the postharvest storage,the β-Gal activity was quite low.

关 键 词：枣 Β-半乳糖苷酶 实时荧光定量PCR 

分 类 号：Q785[生物学—分子生物学]