期刊论文

Prokaryotic expression, polyclonal antibody preparation, expression profiling and functional characterization of the activating enhancer binding protein AP-4 gene from the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

中文

昆虫学报

0454-6296

2019

62

9

1028-1037

10.16380/j.kcxb.2019.09.003

2017YFD0200400:国家重点研发计划 31401762:国家自然科学基金 31672355:国家自然科学基金 2017050304010320:武汉市青年科技晨光计划

本校为第一机构

[目的]激活增强子结合蛋白4 (activating enhancer binding protein 4,AP-4)是近年来备受关注的一类功能广泛的转录因子,参与Wnt/β-catenin信号通路的调控.本研究旨在研究棉铃虫Helicoverpa armigera HaAP-4基因的原核表达并制备多克隆抗体,明确HaAP-4基因的时空表达谱,初步探究HaAP-4对棉铃虫胆固醇载体蛋白2(sterol carrier protein-2,SCP-2)基因(HaSCP-2)表达的调控作用.[方法]通过PCR扩增棉铃虫HaAP-4基因片段,将其克隆至pET-28a原核表达载体,转化至大肠杆菌Escherichia coli BL21,经IPTG诱导表达,采用镍柱纯化重组蛋白,免疫新西兰兔制备多克隆抗体.运用RT-qPCR检测HaAP-4基因在棉铃虫不同...

【Aim】 Activating enhancer binding protein 4 (AP-4), which is involved in the Wnt/β-catenin signaling pathway, is an important transcription factor that has attracted extensive attention in recent years. This study aims to express HaAP-4 gene of Helicoverpa armigera by using prokaryotic expression system, to prepare the polyclonal antibody, to determine the spatio-temporal expression profiles of HaAP-4 and to investigate the possible role of HaAP-4 in regulating the expression of sterol carrier protein-2 gene (HaSCP-2) of H. armigera. 【Metho...