摘要
[目的]本文旨在克隆和研究不结球白菜BcVIL1基因在春化过程中的表达情况,初步了解不结球白菜BcVIL1基因的结构及功能。[方法]以普通白菜PC-175和菜心‘CX-49’为试验材料,利用RT-PCR技术克隆BcVIL1基因全长,并采用生物信息学方法进行氨基酸序列比对和进化分析。通过实时荧光定量PCR(RT-qPCR)技术分析2种材料春化过程中BcVIL1的表达情况。构建亚细胞定位载体pEZS-NL-BcVIL1,利用亚细胞定位技术,分析BcVIL1基因在真核细胞中表达的位置。[结果]2种材料中BcVIL1基因序列基本一致,开放阅读框为1020bp,编码339个氨基酸。进化分析表明:BcVIL1基因与大白菜进化关系最近,同源性最高(99%)。在PC-175中,春化处理7d就可以诱导BcVIL1基因的表达,且在整个春化过程及春化之后都有较高的水平表达,而在‘CX-49’中,BcVIL1基因在春化过程中的表达与未春化处理相比无显著差异。亚细胞定位结果表明:BcVIL1主要在细胞核与细胞膜上表达。[结论]BcVIL1基因在春化过程中其表达具有品种特异性,且在不结球白菜中该基因主要在细胞核与细胞膜上发挥作用。
[Objectives]BcVIL 1 gene had been cloned and some experiments had been producted to better understand the characteristics and structure of BcVIL 1.[Methods]To explore the function of the VIL 1 gene in non-heading Chinese cabbage,PC-175 and‘CX-49’were chosen as materials which belong to typical different genotypes.The BcVIL 1 gene in PC-175 and‘CX-49’was cloned by RT-PCR,and bioinformatics analysis was conducted.The expression patterns of BcVIL 1 in PC-175 and‘CX-49’during the course of vernalization were analyzed by RT-qPCR.The vector pEZS-NL-BcVIL 1 was constructed and the spatial expression of BcVIL 1 was analyzed by sub-cellular localization.[Results]The sequences of BcVIL 1 were consistent in two materials of non-heading Chinese cabbage,and BcVIL 1 containing a 1 020 bp open reading frame encoded 339 amino acid residues.The bioinformatics analysis showed that the homology between Brassica campestris ssp.chinensis Makino and B.rapa ssp.pekinensis was very high at 99%.The BcVIL 1 gene in PC-175 could be induced by 7 days of vernalizaing and the expression of BcVIL 1 gene was at high level during and after the course of vernalization.While the expression of BcVIL 1 gene in‘CX-49’had no obvious differentce compared with non-vernalization condition.The BcVIL 1 gene was verified to be located in cell nucleus and membrane by sub-cellular localization.[Conclusions]The vernalization in PC-175 can induce the expression of BcVIL 1,and the expression of BcVIL 1 also was at high level after vernalization.However the expression of BcVIL 1 gene in‘CX-49’didn’t have this profile.The expression of BcVIL 1 gene was different in two genotypes.BcVIL 1 gene in non-heading Chinese cabbage mainly had a function at cell nucleus and membrane.
作者
申浩冉
肖栋
侯喜林
SHEN Haoran;XIAO Dong;HOU Xilin(State Key Laboratory of Crop Genetics and Germplasm Enhancement/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in Eastern China,Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China)
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2018年第5期825-831,共7页
Journal of Nanjing Agricultural University
基金
国家重点研发计划项目(2016YFD0101701)
关键词
不结球白菜
BcVIL1
克隆
基因表达
亚细胞定位
Brassica campestris ssp.chinensis Makino
BcVIL 1
clone
gene expression
subcellular location
