黄瓜硝酸还原酶基因(CsNR)序列分析及正反义表达载体的构建  

Sequence Analysis and Construction of Sense and Antisense Expressing Vector of Cucumber Nitrate Reductase(CsNR)

作　　者：潘玲玲[1] 王秀峰[1,2] 李强[1] 杨凤娟[1,2] 魏珉[1,3] 史庆华[1,2] PAN Lingling WANG Xiufeng LI Qiang YANG Fengjuan WEI Min SHI Qinghua(College of Horticulture Science and Engineering, Shandong Agricultural University, Tai＇an, Shandong 271018, China State Key Laboratory of Crop Science, Tai＇an, Shandong 271018, China Scientific Observing and Experimental Station of Environment Controlled Agricultural Engineering in Huang-Huai-Hai Region, Ministry of Agriculture, Tai＇an, Shandong 271018, China)

机构地区：[1]山东农业大学园艺科学与工程学院,山东泰安271018 [2]作物生物学国家重点实验室,山东泰安271018 [3]农业部黄淮海设施农业工程科学观测实验站,山东泰安271018

出　　处：《天津农业科学》2017年第7期1-5,共5页Tianjin Agricultural Sciences

基　　金：国家现代农业产业技术体系建设专项资助(CARS-25)

摘　　要：为进一步研究CsNR的作用机制,从黄瓜幼叶中克隆获得硝酸还原酶基因编码区序列(CDS)全长和片段(CsNR;EC1.6.6.1),并运用生物学软件对该基因进行序列分析。其中CDS全长2 748 bp,编码915个氨基酸;CDS片段360 bp。CDS全长和p ROKⅡ经Kpn I单酶切和连接,构建了CsNR正义表达载体;CDS片段和p ROKⅡ经Kpn I和Sac I双酶切,构建了CsNR反义表达载体。通过与其他高等植物NR蛋白进行同源性比对分析,结果发现,CsNR基因的氨基酸序列与甜瓜、烟草、拟南芥、油菜NR基因编码的氨基酸序列高度同源,其中与甜瓜NR蛋白之间同源性最高,为98.25%。To further study the mechanism of CsNR, a fragment and full length CDS of cucumber nitrate reductase（CsNR; EC1.6.6.1） was isolated from cucumber（Cucumis sativus L.） young leaves, respectively. Meanwhile, the gene sequence was analyzed by the biology software. The fragment and full-length CDS was 2 952 and 360 bp, respectively. The full-length CDS and p ROKII were digested to construct sense expressing vector. The fragment CDS and p ROKII were digested to construct sense expressing vector. The homology alignment analysis indicated that the protein encoded by CsNR was in high similarity with NR-encoding protein in Cucumis melo, Nicotiana tabacum, Arabidopsis thaliana and Brassica campestris. The homology with amino acid sequences of CsNR compared with Cucumis melo was the highest by 98.25%.

关 键 词：黄瓜 硝酸还原酶 生物信息学 正义表达载体 反义表达载体 

分 类 号：S642.2[农业科学—蔬菜学]