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本研究运用已经获得的1条490bp的普通烟草钾离子(K~+)通道基因的中间片段来设计特异性引物,应用RACE方法获得其3’和5’末端cDNA序列。通过三段cDNA序列拼接结合全长克隆测序验证,获得一个未报道的普通烟草K~+通道基因,并将其命名为NKT5(NCBI登录号为HM010922)。NKT5的cDNA全长为2365bp,其中5’端非编码区190bp、3’端非编码区249bp、编码区1926bp,编码641AA。对该基因进行了序列分析及功能预测,为进一步阐明其在烟草吸钾及抗逆过程中的功能及分子机制奠定了基础。
In this study, we designed a specific primer using a 490bp intermediate fragment of the tobacco potassium ion (K +) channel gene and obtained its 3 ’and 5’ end cDNA sequences by RACE. An unrecorded gene of K ~ + channel of Nicotiana tabacum was obtained by sequencing the full length of three cDNA segments by sequencing. The gene was named as NKT5 (NCBI accession number HM010922). The full-length cDNA of NKT5 was 2365bp, including 190bp in the 5 ’non-coding region, 249bp in the 3’ non-coding region and 1926bp in the coding region, encoding 641AA. The sequence analysis and function prediction of this gene have laid the foundation for further elucidation of its function and molecular mechanism in the process of potassium uptake and stress resistance in tobacco.