| 翅碱蓬脱水素基因克隆及表达载体构建 |
| |
| 引用本文： | 马慧,李丽丽,祝红艳,钟鸣,陈丽静,郭志富.翅碱蓬脱水素基因克隆及表达载体构建[J].广西农业科学,2013(6):893-897. |
| |
| 作者姓名： | 马慧 李丽丽 祝红艳 钟鸣 陈丽静 郭志富 |
| |
| 作者单位： | 沈阳农业大学生物科学技术学院,辽宁省农业生物技术重点实验室,沈阳110161 |
| |
| 基金项目： | 辽宁省农业生物技术重点实验室基金项目（200556）; 辽宁省农业科学院博士后基金项目（2012） |
| |
| 摘 要： | 【目的】克隆翅碱蓬脱水素蛋白基因并构建其植物表达载体,为其耐盐性研究奠定基础。【方法】提取翅碱蓬总RNA,通过同源克隆和RACE方法分离获得翅碱蓬脱水素蛋白基因,并构建该基因的植物表达载体pBI121-DHN。【结果】克隆获得翅碱蓬脱水素蛋白基因的全长cDNA为847bp（GenBank序列编号：KC013239）,命名为SsDHN。序列分析结果表明,SsDHN全长cDNA中5＇-UTR为61bp,ORF为678bp,3＇-UTR为108bp且包含29bp的poly（A）尾巴,其起始密码子ATG位于62bp处,终止密码子TAA位于737bp处,编码225个氨基酸。氨基酸比对结果表明,该基因推导的氨基酸与已知其他植物DHN基因序列有35%～48%的相似性。聚类分析结果显示,翅碱蓬（SsDHN）与拟南芥和荠菜DHN亲缘关系较近,与咖啡、杜鹃花的亲缘关系较远。【结论】成功克隆了翅碱蓬脱水素基因,并构建了其表达载体,可用于翅碱蓬抗盐的遗传改良。
|
| 关 键 词： | 盐地碱蓬 脱水素蛋白 基因克隆 表达载体构建 |
Cloning and expression vector construction of Suaeda salsa dehydrin gene |
| |
| MA Hui,LI Li-li,ZHU Hong-yan,ZHONG Ming,CHEN Li-jing,GUO Zhi-fu.Cloning and expression vector construction of Suaeda salsa dehydrin gene[J].Guangxi Agricultural Sciences,2013(6):893-897. |
| |
| Authors: | MA Hui LI Li-li ZHU Hong-yan ZHONG Ming CHEN Li-jing GUO Zhi-fu |
| |
| Institution: | (College of Biological Science and Technology,Shenyang Agricultural University,Liaoning Key Laboratory of Agricultural Biotechnology,Shenyang 110161,China) |
| |
| Abstract: | Objective Cloning and expression vector of Suaeda salsa dehydrin gene were conducted to provide references for enduring-salt research of Suaeda salsa. Method Total RNA of Suaeda salsa was extracted, and the gene of Suaeda salsa dehydration was obtained using homology cloning and RACE separation methods. The plant expression vector pBI121-DHN of this gene was constructed and transformed into Agrobacterium tumefaciens for genetic transformation. ResultThe full-length cDNA of Suaeda salsa dehydrin gene was 847 bp （GenBank sequence number： KC013239） and named SsDHN. 5＇-UTR in the full length of SsDHN was 61 bp, and 3＇-UTR zone was 108 bp including a polyA tail with 29 bp. Its ATG, the initiation codon, was located in 62 bp, and TAA, termination codon, was in 737 bp and encoded 225 amino acids. The results of deduced amino aicd comparison indicated that gene sequence of SsDHN had 35%-48% similarity with the other known plant DHN. The cluster analysis results showed that phylogenetic relationship between SsDHN and DHN of Arabidopsis thaliana and Capsella bursa-pastoris was genetically close. However, SsDHN was not closely related to Coffea canephora and Rhododendron catawbiense. Conclusion The dehydrin gene of Suaeda salsa was cloned successfully. The plant expression vector was successfully constructed and can be used for the plant genetic improvement in salt resistance of Suaeda salsa. |
| |
| Keywords: | Suaeda salsa dehydration grain protein gene cloning expression vector construction |
| 本文献已被 维普 等数据库收录！ |
|
