摘要
以‘粤广节瓜’为材料,克隆得到基因CqCHP1(编码富含半胱氨酸和组氨酸C1结构域)。CqCHP1的开放阅读框为816 bp,编码271个氨基酸残基,蛋白二级结构主要由无规则卷曲构成,含23个丝氨酸磷酸化位点。进化树与氨基酸序列比对分析发现CqCHP1和黄瓜Csa_4G297435同源性最高。qRT-PCR分析表明,CqCHP1在节瓜不同组织中均有表达,并受低温显著诱导;苗期和结果期时,CqCHP1在耐冷材料AW66中的表达量显著高于冷敏材料AW9,但在营养生长期和开花期冷敏材料AW9的表达显著高于耐冷材料AW66。亚细胞定位结果显示该蛋白定位于细胞核中。利用酵母文库进行互作蛋白筛选发现CqCHP1可以与bHLH92在细胞核中互作,且双分子荧光互补实验和荧光素酶互补实验验证了其互作的准确性。
In this study,CqCHP1 was cloned from‘Yueguang Chieh-qua’,which encoded cysteine/histidine-rich C1 domain protein.The gene’s open reading frame was 816 bp with 271 amino acids.The secondary structure indicated that CqCHP1 contained main random curl and 23 serine phosphorylation sites.Phylogenetic tree and amino acid sequence alignment analysis showed that CqCHP1 had the highest homology with cucumber Csa_4G297435.CqCHP1 was expressed in different tissues and specifically induced by low temperature.And at seedling and fruiting stages,CqCHP1 showed higher expression in resistance materials than sensitive materials,while at vegetative growth and flowering periods,it’s the opposite.The protein was located in the nucleus.Analysis of interaction proteins obtained by yeast screening showed that CqCHP1 could interact with bHLH92 in nucleus.And the inteaction was verified by Bimolecular fluorescence complementation(BIFC)and Luciferase complementation imaging assay(LCI).
作者
王敏
杨松光
刘微
何晓明
史绍琪
刘文睿
陈林
江彪
彭庆务
WANG Min;YANG Songguang;LIU Wei;HE Xiaoming;SHI Shaoqi;LIU Wenrui;CHEN Lin;JIANG Biao;PENG Qingwu(Guangdong Key Laboratory for New Technology Research of Vegetables,Vegetable Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China)
出处
《园艺学报》
CAS
CSCD
北大核心
2023年第11期2376-2386,共11页
Acta Horticulturae Sinica
基金
国家自然科学基金项目(32002038)
广州市科技计划项目(202102020750)
科技创新战略专项资金(高水平农科院建设)项目(R2021PY-QF008)
广东省农业科学院农业优势产业学科团队建设项目(202103TD,202114TD)。
作者简介
通信作者:彭庆务,E-mail:pengqingwu@gdaas.cn。
