摘要
目的:从星星草中克隆一个铁蛋白相关基因,分析其序列特征及基因表达模式。方法与结果:构建星星草RACE cDNA文库,根据GenBank中报道的铁蛋白基因EST序列设计引物,利用RACE方法克隆得到星星草铁蛋白基因PtFer的全长cDNA序列(GenBank登录号为HM125047);序列分析表明,PtFer的核苷酸序列长度为751 bp,开放读框为336 bp,编码111个氨基酸残基,预测蛋白质相对分子质量为12.8×103。其蛋白质序列具有铁蛋白的特征性保守区域,与水稻属同一进化分支,与其他禾本科植物铁蛋白的序列相似性达80%以上。Northern杂交分析表明,PtFer的表达量随Na2CO3的浓度和时间的增加而升高,并在12～24 h内持续保持较高的表达水平。结论:用分子生物学及生物信息学技术克隆并分析了星星草铁蛋白基因PtFer的全长cDNA序列及编码蛋白的结构,并初步探索了盐碱胁迫下其表达模式,为研究植物耐盐碱机理奠定了基础。
Objective：To clone a ferritin related gene of Puccinellia tenuiflora and analyze the sequence characteristics and gene expression patterns.Methods ＆ Results：The RACE cDNA library was constructed,and the primers were designed in terms of the EST in GenBank.A ferritin gene PtFer（GenBank accession number is HM125047） was cloned from the cDNA library of P.tenuiflora by RACE technique.Sequence analysis revealed that the full length of PtFer cDNA was 751 bp,including 336 bp open reading frame,which encoding 111 amino acid,and the predicted protein molecular mass was 12.8 kD.The predicted protein sequence of PtFer had the typical ferritin conserved domain.It shared the same evolutionary branch with rice and there was more than 80% sequence identity to other gramineae plants’ ferritin.Northern blotting analysis showed that the expression level of PtFer was increased with the Na2CO3 concentration and time gradient and kept most between 12～24 h.Conclusion：The gene PtFer of P.tenuiflora was cloned,and its cDNA sequence,structural feature of encoding protein and the expression levels were analyzed by molecular biology and bioinformatics techniques.All of above can provide important information regarding the salt-resistant mechanism of P.tenuiflora
出处
《生物技术通讯》
CAS
2011年第1期32-36,共5页
Letters in Biotechnology
基金
国家高技术研究发展计划(2006AA10Z129
2008AA10Z156)
黑龙江省科技计划(攻关)项目(GB06B112-5)
关键词
星星草
铁蛋白
基因克隆
表达分析
盐碱胁迫
Puccinellia tenuiflora
ferritin
alkali-stress
gene cloning
expression analysis
