文献类型：期刊文献

中文题名：杨树MADS转录因子SVL基因克隆、原核表达与多克隆抗体制备

英文题名：Cloning,Prokaryotic Expression and Polyclonal Antibody Preparation of Populus MADS Transcriptional Factor Members PtSVL

作者：王冬莉[1] 刘芸杉[1,2] 孟森[1] 卞展[1]

第一作者：王冬莉

机构：[1]中国林业科学研究院热带林业研究所,广东广州510520;[2]重庆三峡学院生物与食品工程学院,重庆404100

年份：2022

卷号：37

期号：3

起止页码：82-87

中文期刊名：西北林学院学报

外文期刊名：Journal of Northwest Forestry University

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2021_2022】；

基金：中国林业科学研究院青年协同创新项目(CAFYBB2019QD001-4)。

语种：中文

中文关键词：杨树;MADS转录因子;PtSVL;原核表达;多克隆抗体

外文关键词：Populus;MADS transcriptional factor;PtSVL;prokaryotic expression;polyclonal antibody

分类号：S792.11

摘要：为研究杨树MADS转录因子相关基因在逆境中的作用,以84K杨树叶片的cDNA为模板,通过PCR扩增得到了开放阅读框为684 bp、氨基酸个数为227的PtSVL基因;构建原核表达载体pET-B2M-PtSVL-3×FLAG,并在大肠杆菌B21(DH3)中大量表达。结果表明,获得的PtSVL-3×FLAG融合蛋白,大小为42.0 ku,主要以包涵体形式存在;用纯化后的蛋白免疫新西兰公兔制备PtSVL多克隆抗体;间接ELISA法和Western blot检测后多克隆抗体后显示其效价高,特异性好。获得的PtSVL多克隆抗体将为后续鉴定PtSVL蛋白的互作蛋白提供基础。
In order to understand the role of MADS transcriptional factor in response to adversity stresses,PtSVL gene was obtained from the cDNA of 84K(Populus alba×P.glandulosa'84K')leaves using PCR amplification with he full length of 684 bp,encoding 227 amino acids.The recombinant vector pET-B2M-PtSVL-3×FLAG was expressed in E.coli strain B21(DH3).The fusion PtSVL and three FLAG tags protein was 42.0 ku and mainly existed in inclusion body.By immunizing the rabbits with the purified protein,polyclonal antibody of PtSVL was produced.ELISA and Western blot showed that the antibody had high titer and good specificity.The obtained antibody can be used for subsequent PtSVL protein interactions.