[1]王冬莉,刘芸杉,孟森,等. 杨树MADS转录因子SVL基因克隆、原核表达与多克隆抗体制备[J].西北林学院学报,2022,37(3):82-87.[doi:10.3969/j.issn.1001-7461.2022.03.12]
 WANG Dong-li,LIU Yun-shan,MENG Sen,et al. Cloning,Prokaryotic Expression and Polyclonal Antibody Preparation of Populus MADS Transcriptional Factor Members PtSVL[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2022,37(3):82-87.[doi:10.3969/j.issn.1001-7461.2022.03.12]
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 杨树MADS转录因子SVL基因克隆、原核表达与多克隆抗体制备()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第37卷
期数:
2022年第3期
页码:
82-87
栏目:
出版日期:
2022-05-31

文章信息/Info

Title:
 Cloning,Prokaryotic Expression and Polyclonal Antibody Preparation of Populus MADS Transcriptional Factor Members PtSVL
文章编号:
1001-7461(2022)03-0082-06
作者:
 王冬莉1刘芸杉12孟森1卞展1*
 (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)
Author(s):
 WANG Dong-li1LIU Yun-shan12MENG Sen1BIAN-Zhan1*
 (1.Research Institute of Tropical Forestry,Chinese Academy of Forestry,Guangzhou 510520,Guangdong,China; 2.College of Biology and Food Engineering,Chongqing Three Gorges University,Chongqing 404100,China)
关键词:
 杨树MADS转录因子PtSVL原核表达多克隆抗体
Keywords:
Populus MADS transcriptional factor PtSVL prokaryotic expression polyclonal antibody
分类号:
S792.11
DOI:
10.3969/j.issn.1001-7461.2022.03.12
文献标志码:
A
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。
Abstract:
 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.

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 LIU Dan,LU Hua-tao,JIANG Yue-zhong,et al. Evaluation of Resistance to Cerambycidae of Poplar Clones[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2012,27(3):72.
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备注/Memo

备注/Memo:
 收稿日期:2021-06-01修回日期:2021-07-01
基金项目:中国林业科学研究院青年协同创新项目(CAFYBB2019QD001-4)。
第一作者:王冬莉。研究方向:植物分子生物学研究。E-mail:wangdongli1997@163.com
*通信作者:卞展,助理研究员。研究方向:植物分子生物学研究。E-mail:bianzhanzhan@163.com
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