红麻液泡膜质子泵H^+-PPase(Hcvp1)基因的克隆、序列分析和表达  

Cloning and Analysis of Sequence and Expression of a Vaculoar H^+-PPase Gene in Kenaf

作　　者：李辉[1] 李德芳[1] 陈安国[1] 唐慧娟[1] 李建军[1] 黄思齐[1] 

机构地区：[1]中国农业科学院麻类研究所,湖南长沙410205

出　　处：《华北农学报》2017年第1期9-14,共6页Acta Agriculturae Boreali-Sinica

基　　金：基金项目 国家麻类产业技术体系红麻育种岗位(CARS-19-E07);中国农业科学院科技创新工程(ASTIP-IBFC03)

摘　　要：为了阐明红麻耐盐机理,以耐盐性较强的中红麻16号为材料,从红麻耐盐转录组测序结果中获得了1个与AVP1基因高度相似的Unigene,根据该片段的序列设计引物,直接进行PCR扩增,经Sanger测序获得该基因全长c DNA序列。对该基因序列进行生物信息学分析表明:该基因全长2 298 bp,开放阅读框为2 298 bp,编码765个氨基酸,其编码蛋白质的分子量和等电点分别为80.2 k Da和5.25,与雷蒙德氏棉、甜橙、毛果杨、烟草和大豆的H+-PPase基因核苷酸序列的相似性分别为90%,85%,85%,83%,83%,蛋白质序列的相似性分别为96%,93%,91%,93%,94%,说明该基因与AVP1为同源基因,命名为Hcvp1。qRT-PCR分析表明,该基因的表达量随着Na Cl浓度的增加而增加。这将为Hcvp1基因的功能验证和红麻耐盐机理的研究奠定坚实的基础。In order to elucidate the mechanism of salt tolerance,taking Zhonghongma 16 with salinity tolerance as an experimental material,we obtained a kenaf Unigene highly identified with AVP1 gene from the kenaf transcriptome sequence. According to the known fragment,we developed primer to amplify directly by PCR,and the c DNA sequence of the gene was obtained by Sanger sequencing. The results of bioinformatics analysis showed that the full length of the Hcvp1 gene was 2 298 bp,the open reading frame was 2 298 bp,coding 765 amino acids,the molecular weight and the isoelectric point of coded protein were 80. 2 k Da and 5. 25. Nucleotide sequence Blast indicated that the Unigene of kenaf shared an identity of was 90%,85%,85%,83%,83%,with AVP1 of Gossypium raimondii,Citrus sinensis,Populus trichocarpa,Nicotiana tabacum,Glycine max,respectively,and the similarity in protein Blast was 96%,93%,91%,93%,94% respectively. The results showed that it was a homologous gene of AVP1 in kenaf and named Hcvp1. qRT-PCR analysis indicated that the expression level of Hcvp1 went up while the rise of Na Cl concentration. This study will lay a solid foundation for the Hcvp1 gene function validation and the research of salt tolerance mechanism in kenaf.

关 键 词：红麻 H^+-PPase基因 耐盐 

分 类 号：Q78[生物学—分子生物学]