摘要
对豌豆蚜Jun蛋白进行原核表达进而进行抗体制备,可为探究JNK通路对靶基因的调控机制奠定材料基础。本研究提取豌豆蚜的总RNA,经反转录、PCR扩增后进行原核表达与抗体制备,并用ELASA法测定其抗血清效价、Western-Blot法检测抗体特异性。结果表明:本文所构建的pET-28a-SUMO表达载体能在大肠杆菌体内成功表达出Jun蛋白,并通过免疫兔子成功制备了多克隆抗体,其抗血清效价良好、抗体特异性较好。本研究成功表达出豌豆蚜Jun蛋白,并成功制备出抗体,可用于下步试验,对进一步探究JNK通路对靶基因的调控机制具有重要意义。
Prokaryotic expression of Jun protein from pea aphid,followed by antibody preparation,can lay a mate-rial foundation for exploring the regulatory mechanism of JNK pathway on target genes.In this study,total RNA of pea aphid was extracted,and then prokaryotic expression and antibody preparation were performed after reverse transcrip-tion and PCR amplification,and its antiserum potency was determined by ELASA method and antibody specificity was detected by Western-Blot method.The results showed that the pET-28a-SUMO expression vector constructed in this paper could successfully express Jun protein in Escherichia coli,and successfully prepare polyclonal antibodies by immunizing rabbits,the antibodies had good antiserum potency and good antibody specificity.In this study,we success-fully expressed the Jun protein of pea aphid and prepared antibody that can be used in the next experiments,which is important to further investigate the regulatory mechanism of JNK pathway on target genes.
作者
苏慧迪
王文涛
刘彦
马力
SU Huidi;WANG Wentao;LIU Yan;MA Li(College of Plant Protection,Shanxi Agricultural University,Jinzhong Shanxi 030801)
出处
《现代农业科技》
2023年第22期60-64,共5页
Modern Agricultural Science and Technology
基金
山西省基础研究计划-青年科学研究项目(202103021223125)
2022年山西省高等学校大学生创新创业训练项目(20220157)。
关键词
豌豆蚜
Jun蛋白
原核表达
抗体制备
pea aphid
Jun protein
prokaryotic expression
antibody preparation
