烟草渗调蛋白基因的分离克隆及序列分析  

Cloning and Sequencing of the Gene Encoding Osmotin From Nicotiana Tabacum

作　　者：鄢波[1] 陈莉[1] 张绍松[1] 黄兴奇[1] 

机构地区：[1]云南省农科院生物技术研究所

出　　处：《生物学杂志》1997年第6期10-12,共3页Journal of Biology

基　　金：云南省自然科学基金

摘　　要：参照国外报道的渗调蛋白基因序列，自行设计并合成了一对寡核苷酸引物，通过ＰＣＲ技术从云南地方栽培的烤烟品种“红花大金元”基因组ＤＮＡ中直接扩增出了其全编码序列。ＰＣＲ产物纯化后直接克隆到ｐＧＥＭ－Ｔ载体系统中，转化大肠杆菌ＤＨ５α，在Ｘｇａｌ平板上筛选白色菌落，经ＳｐｈＩ和ＳａｃＩ双酶切，鉴定所得的重组质粒中含有７５３ｂｐ左右的ＰＣＲ扩增片段。采用双链双脱氧法定序分析表明所克隆的渗调蛋白基因编码区序列与国外迄今所报道的两个株系的序列完全一致。According to the nucleotide sequence of the Nicotiana tabacum osmotin gene that reported abroad,we designed and synthesized a pair of primer.DNA was isolated from the tobacco (nicotiana tabacum cv.Hong hua da jin yuan) that was cultivared in Yunnan province.A 0.75 Kb DNA fragment was obtained with PCR technique.After purification,the DNA fragment encoding osmotin was inserted into pGEM-T vector system directly and fransformed into E.coli DH5α cells.The white clone named pOSY6 from screening with Xgal was chosen for further analysis of restriction map by Sph Ⅰ/Sac Ⅰ digestion.The results indicate that the recombinant plasmid pOSY6contains a 0.75Kb amplified DNA fragment.The DNA sequence and deduced amino acid sequence reported here was compared with that published abroad and no difference was found.It is implying high conservation in evolution.

关 键 词：烟草 渗调蛋白 基因克隆 序列分析 

分 类 号：S572.032[农业科学—烟草工业]