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当前位置: 首页 > 文章 > 普通烟草K~+通道基因NKT4的克隆、序列和表达分析 植物遗传资源学报 2009,10 (3) 354-359
Position: Home > Articles > Cloning and Sequence and Expression Analysis of Potassium Channel Gene NKT4 in Nicotiana tabacum Journal of Plant Genetic Resources 2009,10 (3) 354-359

普通烟草K~+通道基因NKT4的克隆、序列和表达分析

作  者：
曲平治;刘贯山;刘好宝;司丛丛;胡晓明;冯祥国;刘朝科
单  位：
川渝中烟工业公司;中国农业科学院烟草研究所
关键词：
普通烟草;K+通道基因;克隆;RACE;表达
摘  要：
通过比对拟南芥、胡萝卜、番茄和马铃薯的K+通道氨基酸序列得到了保守序列,设计1对简并引物,利用RT-PCR获得3条490bp的普通烟草K+通道基因中间片段。以其中一条中间片段设计特异性引物,应用RACE方法得到5′末端和3′末端cDNA序列。通过拼接并结合全长克隆及测序验证,获得一个未报道的普通烟草K+通道基因,并将其命名为NKT4(Gen-Bank登录号为FJ233071)。NKT4的cDNA全长为2937bp,其中5′非编码区45bp、编码区2679bp、3′非编码区213bp;编码区编码892个AA。构建了一个烟草、拟南芥及相关植物K+通道蛋白的系统进化树。基因表达分析表明,NKT4主要在烟草主根和侧根中表达,在烟草叶中也有少量表达。
译  名：
Cloning and Sequence and Expression Analysis of Potassium Channel Gene NKT4 in Nicotiana tabacum
作  者：
QU Ping-zhi1,LIU Guan-shan1,LIU Hao-bao1,SI Cong-cong1,HU Xiao-ming2,FENG Xiang-guo2,LIU Chao-ke2(1Tobacco Research Institute,Chinese Academy of Agricultural Sciences,Qingdao 266101;2China Tobacco Chuanyu Industrial Corporation,Chengdu 610000)
关键词：
Nicotiana tabacum;K+ channel gene;Cloning;RACE;Expression
摘  要：
A pair of degenerate primers was designed based on the conserved amino-acid sequence that was got by the alignment of K+ channel protein sequences from Arabidopsis thaliana,carrot,tomato and potato,then three strips of 490 bp segments were first isolated by RT-PCR.5′ and 3′-ends of cDNA were acquired by RACE strategy using the specific primers that were designed according to one of the segment cDNA.A unreported Nicotiana tabacum K+ channel gene was obtained by splicing and the identification of full-length cloning.The cDNA sequence has been submitted to the NCBI GenBank,named as NKT4 and the accession number is FJ233071.The full-length cDNA of NKT4 is 2937 bp long,with 45 bp 5′-untranslated region,2679 bp coding sequence and 213 bp 3′-untranslated region.The deduced amino acid is 892 AA long.A phylogenetic tree of K+ channels from Nicotiana tabacum,Arabidopsis thaliana and related plants was constructed.NKT4 was mainly expressed in main roots and lateral roots and little in leaves in Nicotiana tabacum.

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