摘要
提取新牧一号杂花苜蓿的总RNA,用特异引物RT-PCR扩增后的cDNA片段连接入pMD19-T载体,转化大肠杆菌DH5α,对阳性克隆进行序列分析,新牧一号苜蓿NHX1(MvNHX1)全长1 626 bp,Genbank登录号为:EU375310。半定量RT-PCR和实时荧光定量PCR分析表明,在盐胁迫下MvNHX1基因的表达均上调。构建重组植物表达载体pBI121-MvNHX1后,通过农杆菌介导的叶盘法转化烟草,转基因烟草在盐胁迫下发芽率和生物量均高于非转基因烟草,表明MvNHX1能够提高转基因烟草的耐盐性。
The total RNA of Medicago varia xinmu-1 was extracted,the cDNA fragment amplified by RT-PCR using special primer was linked into pMD19-T vector and transformed into Escherichia coli DH5α.By sequencing the positive clone,the MvNHX1（EU375310） is 1626bp long.RT-PCR and real-time PCR assays showed that the level of MvNHX1 transcription was up-regulated and reached a steady higher level in the seedlings after high salinity treatment.The MvNHX1 gene was transformed into tobacco via agrobacterium mediation after the plant expression vectors pBI121-MvNHX1 was successfully constructed.The germination rate and biomass of the transgenic tobacco were taller than those of the control group under NaCl stress,indicating that the MvNHX1 can enhance the salt tolerance of transgenic tobacco.
出处
《植物研究》
CAS
CSCD
北大核心
2011年第5期550-557,共8页
Bulletin of Botanical Research
基金
国家转基因重大专项2009ZX08005-022B、2008ZX08005-004(4-3)
