番茄LeNHX1基因植物过量表达载体的构建及表达分析

Expression Analysis and Construction of Plant Over-expression Vector of Tomato LeNHX1 Gene

从番茄幼苗中提取RNA,根据NCBI中番茄LeNHX1基因序列设计引物,通过RT-PCR获得了番茄LeNHX1基因的cDNA序列,包含一个1 605 bp的开放阅读框,编码534个氨基酸。将cDNA序列连接到植物过量表达载体PBI121上,对所获得的重组质粒进行双酶切鉴定,结果表明,植物过量表达载体PBI121-LeNHX1已构建成功。半定量RT-PCR结果表明LeNHX1基因在根、茎和叶中均表达,盐、低温和脱落酸的诱导能提高LeNHX1基因的表达量,推测番茄LeNHX1基因在逆境应答中可能起着重要作用。

Total RNA was extracted from the tomato young seedling,cDNA was obtained by reverse transcription polymerase chain reaction. Primers were designed according to tomato LeNHX1 gene sequence of NCBl,then the target LeNHX1 gene was amplified by RT-PCR protocol, the nucleotides fragment of LeNHX1 gene including an ORF（ 1605 bp）, encoding 534 amino acids,The transcript of which was connected to plant over-expression vector PBI121. The double digestion of recombinant plasmid result showed that the plant over-expression vectors with LeNHXI was constructed successfully. The results of RT-PCR showed that LeNHX1 was expressed in roots,stems and leaves,the expression level was induced by salt,low temperature and ABA stress. We suggest the LeNHX1 gene could play an important role of stress response in tomato.