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茶树AMP脱氨酶基因的克隆与序列分析
魏艳丽,李金,庞磊,江昌俊
0
(安徽农业大学教育部/农业部茶叶生物化学与生物技术重点开放实验室,安徽农业大学茶与食品科技学院,合肥 230036)
摘要:
茶树AMP脱氨酶是茶树咖啡碱合成途径的关键酶,它的活性影响着咖啡碱的的合成,但该酶基因的克隆目前还处于空白状态。通过从茶树全器官转录组文库搜寻的序列进行比对,获得1条与其他物种同源性较高的编码AMP脱氨酶基因的EST序列,运用RACE技术扩增获得茶树AMP脱氨酶基因的cDNA全长序列。该基因cDNA全长3 215 bp,其中开放阅读框长2 571 bp,编码856个氨基酸,3′端有一个明显的多聚腺苷酸加尾信号。预测蛋白分子量为97.6 kDa,理论等电点为6.31。序列分析表明它与葡萄的AMP脱氨酶基因的亲缘关系较近。将基因登陆到GenBank上,登录号为AGJ84350.1。
关键词:  茶树  AMP脱氨酶  RACE  序列分析
DOI:
投稿时间:2013-04-19
基金项目:国家“十二五”科技支撑计划课题(2011BAD01B01)资助。
Cloning and sequence analysis of AMP deaminase gene from tea plant
WEI Yan-li,LI Jin,PANG Lei,JIANG Chang-jun
(Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education, Ministry of Agriculture, School of Tea and Food, Anhui Agricultural University, Hefei 230036)
Abstract:
AMP deaminase is a key enzyme in tea caffeine synthesis pathway, and its activity affects the synthesis of caffeine, but the AMP deaminase gene of tea plant has not been cloned. We screened an EST from the whole organic transcriptomic library of tea, which had high homology with AMP deaminase gene from other organisms, and amplified through RACE technology to obtain the cDNA full-length of AMP deaminase gene. The cDNA full-length was 3 215 bp with a single 2 571 bp opening reading frame that was predicted to encode 856 amino acids, and its 3′ untranslated region had an obvious polyadenylation signal. The deduced protein molecular weight was 97.6 kDa and its theoretical isoelectric point was 6.31. Sequence analysis result showed that it is closely related with that of Vitis vinifera. The gene was submitted to GenBank and the accession number is AGJ84350.1.
Key words:  tea plant  AMP deaminase  RACE  sequence analysis

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