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当前位置: 首页 > 文章 > 日本结缕草ZjNOL基因的克隆及转录自激活检测 中国草地学报 2019 (3)
Position: Home > Articles > 日本结缕草ZjNOL基因的克隆及转录自激活检测 Chinese Journal of Grassland 2019 (3)

日本结缕草ZjNOL基因的克隆及转录自激活检测

作  者：
董笛;张宇昕;刘畅;王焕英;李殷睿智;刘卓成;王梦迪;晁跃辉;韩烈保
关键词：
日本结缕草;NOL基因;转录自激活;生物信息学
摘  要：
NYC1-like(NOL)编码的短链氧化还原酶是叶绿素b降解的必需酶.对日本结缕草(Zoysia japonica)ZjNOL基因全长克隆结果显示,该基因开放阅读框为981个核苷酸,编码327个氨基酸.生物学分析显示,ZjNOL蛋白与高粱(Sorghum bicolor)中的NOL蛋白的亲缘关系最近.利用pGBKT7载体,通过DNA重组技术成功构建pGBKT7-ZjNOL酵母表达载体,通过PEG介导的转化方法,成功导入Y2HGold酵母中,自激活检测显示含有pGBKT7-ZjNOL的Y2HGold酵母细胞能够在SD/-Trp培养基上正常生长,而在SD/-Trp/AbA培养基上不能生长,也不能在含有X-α-Gal培养基显蓝色,证明ZjNOL蛋白没有自激活活性.生物信息学分析及转录自激活检测有助于深入研究ZjNOL基因功能,为延长结缕草绿期,改善结缕草坪用质量奠定基础.
作  者：
DONG Di;ZHANG Yu-xin;LIU Chang;WANG Huan-ying;LI Yinrui-zhi;LIU Zhuo-cheng;WANG Meng-di;CHAO Yue-hui;HAN Lie-bao;College of Grassland Science of Beijing Forestry University;Baotou Municipal Environmental Protection Bureau Kun District Branch;Beijing Research & Development Center for Grass and Environment;
单  位：
DONG Di%ZHANG Yu-xin%LIU Chang%WANG Huan-ying%LI Yinrui-zhi%LIU Zhuo-cheng%WANG Meng-di%CHAO Yue-hui%HAN Lie-bao%College of Grassland Science of Beijing Forestry University%Baotou Municipal Environmental Protection Bureau Kun District Branch%Beijing Research & Development Center for Grass and Environment
关键词：
Zoysia japonica;;ZjNOL;;Autonomous transcriptional activation;;Bioinformatics analysis
摘  要：
The short-chain dehydrogenase/reductase encoded by NYC1-like(NOL) is an essential enzyme for the degradation of chlorophyll b. The full-length gene clone of ZjNOL in zoysia(Zoysia japonica) showed that the open reading frame consisted of 981 nucleotides and 327 amino acids were encoded by ZjNOL. Biological analysis exhibited that ZjNOL is closely related to NOL protein in sorghum(Sorghum bicolor). The pGBKT7-ZjNOL vector was constructed by DNA recombination technology and successfully introduced into Y2HGold yeast by PEG-mediated transformation method. Autonomous transcriptional activation testing showed that Y2HGold yeast cells containing pGBKT7-ZjNOL can be cultured in SD/-Trp,but not on SD/-Trp/AbA medium. Furthermore, it cannot show blue color in the medium containing X-α-Gal. All above results demonstrated that ZjNOL protein had no self-activation activity. Bioinformatics analysis and autonomous transcriptional activation testing can facilitate further study on the function of ZjNOL gene, which provide basis for extending the green period and improving the quality of zoysia turfgrass.

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