茶树GAGP基因克隆及表达分析

Gene Cloning and Expression Analysis of GAGP in Tea Plant

采用SSH技术分析了VA菌根处理后福鼎大白茶根系基因差异表达情况,获得了差异序列,序列比对显示,在下调表达序列中可能包含了10种未知功能的基因;在上调表达序列中可能包含了5种可能的基因。采用RACE技术获得GAGP基因长3 146 bp(Gen Bank,登录号KJ946251),具有2 769 bp开放阅读框(1st^2 769th),编码923个氨基酸。分子生物信息学分析表明,GAGP蛋白分子量约106.9 k D,等电点为8.42,定位于线粒体内。定量PCR分析表明,GAGP在茶树叶片中表达强度存在明显的品种差异,GAGP对生物性和非生物性胁迫均有明显响应。

By using SSH, the differences in gene expression of root from Fudingdabai tea plant infected by VAmycorrhiza were analyzed and the diversity sequences was obtained. The sequence alighment showed that thedown-regulated expression sequence possibly contained 10 unknown genes and the up-regulated expression sequencepossibly contained 5 known genes. The GAGP （gap-pol） genic full-length sequence was obtained by using RACE.The length of GAGP gene was 3146bp （GenBank, Accession no., KJ946251）, with 2 769 bp ORF （1^st-2 769^th）, thesequence encoded 923 amino acid. Bioinformatics indicated that the GAGP protein’s molecular weight was about106.9 kD, IEP was 8.42, located in mitochondria. The study also showed expression degree of GAGP was distinct indifferent cultivars, while it responded obviously to biological and non-biological stress.