摘要
利用RACE(rapid-amplification of cDNA end)方法,以大豆叶片提取总的RNA为模板克隆了大豆Na+/H+逆向转运蛋白(Glycine max Na+/H+antiporter,GmNHX)基因,并将其连接到表达载体PBI121中,构建重组表达载体PBI121-NHX3.分析结果表明:该基因的ORF为1 503 bp,推测编码501个氨基酸.与所选取的10种植物同类蛋白氨基酸序列进行对比,一致性为72%～94%,并具有真核生物单价阳离子(氢离子)反向转运蛋白典型的结构域,将该基因命名为GmNHX3,GenBank接收号为JN872904.通过PCR和酶切鉴定,PBI121-NHX3构建成功.
RACE（rapid-amplification of cDNAends） technique was used to clone Na＋/H＋ antiporter gene from Glycine max L Merr with the total RNA as the template.This gene was linked to the plasmid PBI121 and the expression vector PBI121-NHX3 was constructed.The results show its ORF（open reading frame）consists of 1 503 bp,which can be deduced to encode 501 amino acids.It was found that predicted amino acid sequence had an identification of 72%-94% compared with amino acid sequence from other ten plant species and it had a typical structural domain of the monovalent cation（proton） antiporter.The gene was named GmNHX3,（GenBank accession number： JN872904）.Genetic transformation vector of PBI121-NHX3 was constructed successfully by identification method of PCR and restriction enzyme digestion.
出处
《吉林大学学报（理学版）》
CAS
CSCD
北大核心
2012年第2期365-370,共6页
Journal of Jilin University:Science Edition
基金
内蒙古自然科学基金(批准号:2010BS0502)
内蒙古自治区高等学校研究项目(批准号:NJ10111)
