斑茅-δOAT基因克隆及其序列分析  

Cloning and Sequence Anaylisis of δ-OAT Gene from Erianthus arundinaceus

作　　者：吴杨[1,2] 贺俐[2] 李伟[1] 张木清[1] 

机构地区：[1]福建农林大学农业部甘蔗生理生态与遗传改良重点实验室,福州350002 [2]井冈山大学生命科学学院,吉安343009

出　　处：《植物研究》2009年第5期577-584,共8页Bulletin of Botanical Research

基　　金：国家"863"计划课题糖料新品种选育课题(2004AA241191);福建省自然科学基金计划资助项目(2006J0003)

摘　　要：利用RT-PCR和RACE技术从斑茅(Erianthus arundinaceus)中分离出编码鸟氨酸-δ-氨基转氨酶基因的全长cDNA序列,序列全长1 680 bp,编码454个氨基酸。通过对哺乳动物、高等植物、微生物的-δOAT基因编码的氨基酸序列进行同源比对,发现斑茅-δOAT基因同其近缘属植物甘蔗的同源性最高(87%),同其他高等植物的同源性次之(约为70%),而同动物的同源性最低(约为60%)。在斑茅-δOAT基因编码的氨基酸序列的5′端未发现线粒体定位序列,同甘蔗-δOAT基因一样。斑茅-δOAT基因具有完整的鸟氨酸转氨酶功能区rocD。利用定量RCR(real-tim e PCR)对30%PEG胁迫下的斑茅-δOAT基因表达量进行研究,结果表明-δOAT基因在胁迫12 h表达量达到最高,约为对照的4.1倍;胁迫2 h-δOAT基因表达量反而有所降低。The complete cDNA sequence of orn-δ-aminotransferase （OAT） gene was obtained from Erianthus arundinaceus and was cloned by reverse-transcript-polymerase-chain-reaction （RT-PCR） and rapid amplification of cDNA end （RACE） technologies. The acquired gene was 1680 bp in full length, encoding 454 amino acid residues. The amino acid sequence blast results showed that compared to that from mammal, higher plant and microorganism, δ-OAT gene from E. arundinaceus shared the highest homology （87%） with relative genera plant, Saccharum officinarurn, 70% homology with other higher plants and 60% homology with animal. No N-terminal mitochondrial transit peptide （MTP） was found in the amino acid sequence encoded by δ-OAT gene from E. arundinaceus, which was the same as that from S. officinarum. Complete domain of OAT, rocD, was included in δ-OAT gene from E. arundinaceus. Expression level of δ-OAT gene from E. arundinaceus treated with 30% polyethylene glycol （PEG） was studied using real-time PCR technology, which showed that after treated 12 hours with PEG, the expression level reached the highest, 4.1 times as that of the comparison, but got lower after stressed 2 hours.

关 键 词：斑茅 鸟氨酸-δ-氨基转氨酶 鸟氨酸途径 定量PCR 

分 类 号：Q78[生物学—分子生物学]