百合水通道蛋白LotNIP5-1基因的克隆及表达分析

发布时间：2018-03-17 01:31

本文选题：百合　切入点：NIP-基因克隆　出处：《中国农业大学学报》2017年03期 　论文类型：期刊论文

【摘要】：为研究NIP5-1基因(Nodulin26-like intrinsic protein)在百合生长发育方面的作用机理,利用RACE技术克隆得到百合NIP5-1基因的全长cDNA序列。运用多个软件进行生物信息学分析;采用实时荧光定量PCR(qRT-PCR)及RT-PCR技术检测其表达特性。结果表明:1)百合NIP5-1基因全长为1 492bp,包含1个900bp的完整开放阅读框(Open reading frame,ORF),共编码299个氨基酸;2)百合NIP5-1蛋白属于跨膜通道蛋白MIP超家族。具有6个跨膜区;在B、E环分别具有NIP5-1蛋白特有的NPS、NPV模体,具有该蛋白Ar/R filter分子筛特有的AIGR模体;与小果野芭蕉、香瓜、葡萄、黄瓜、番茄的相似性分别为86%、83%、84%、83%和82%;3)荧光定量结果显示,百合NIP5-1基因在茎中表达量最高,在花瓣中表达量较低,在叶、鳞茎、根中表达量均极低。RTPCR半定量检测显示,其在"湿"柱头上表达量显著高于"干"柱头,这与转录组测序结果相符合。上述研究暗示,百合NIP5-1可能是硼酸通道蛋白,在促进茎生长及生殖活动中发挥作用。
[Abstract]:In order to study the mechanism of NIP5-1 gene Nodulin 26-like intrinsic protein in lily growth and development, the full-length cDNA sequence of lily NIP5-1 gene was cloned by RACE technique. The expression characteristics of NIP5-1 gene were detected by real-time fluorescence quantitative PCR and RT-PCR. The results showed that the total length of NIP5-1 gene was 1492 BP, containing a 900bp complete open reading frame, encoding 299 amino acids and 2 amino acids. The NIP5-1 protein of lily was transmembrane. Channel protein MIP superfamily with 6 transmembrane regions; The NIP5-1 motifs of NIP5-1 protein and AIGR motifs specific to Ar/R filter molecular sieve were found in the E ring, and the similarity with small fruit field plantain, cantaloupe, grape, cucumber and tomato were 883% and 822%, respectively. Lily NIP5-1 gene expression was the highest in stem and low in petal. RT-PCR semi-quantitative analysis showed that the expression of Lilium NIP5-1 gene in "wet" stigma was significantly higher than that in "dry" stigma. These results suggest that NIP5-1 may be a borate channel protein and play an important role in promoting stem growth and reproductive activity.
【作者单位】：北京农学院园林学院;
【基金】：北京市科技提升计划(TJSHG201310020020) 北京林果业生态环境功能提升协同创新中心(PXM2016-014207-000038) 北京市属高等学校创新团队建设项目(IDHT20150503) 城乡生态环境北京实验室项目(PXM2016-014207-000003)
【分类号】：S682.29

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