毛果杨漆酶基因PtrLAC-2的原核表达与多克隆抗体制备研究
Botanical Research 植物学研究 , 2018, 7(2), 177-185 Published Online March 2018 in Hans.
文章引用 : 李盈 , 李嘉鑫 , 滕晓瞳 , 郭晓蕊 , 程子义 , 尹玢 , 张永卓 , 陆海 , 李慧 . 毛果杨漆酶基因 PtrLAC -2的原核表 达与多克隆抗体制备研究 [J]. 植物学研究 , 2018, 7(2): 177-185. DOI: 10.12677/br.2018.72024
Prokaryotic Expression and Polyclonal Antibody Preparation of PtrLAC -2 Gene of Populus trichocarpa
Ying Li, Jiaxin Li, Xiaotong Teng, Xiaorui Guo, Ziyi Cheng, Bin Yin, Yongzhuo Zhang, Hai Lu, Hui Li
College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing
Received: Mar. 2nd , 2018; accepted: Mar. 21st , 2018; published: Mar. 28th , 2018
Abstract
Objective: To clone and express the Populus trichocarpa laccase PtrLAC-2 gene. This study pre-pared polyclonal antibody with high affinity and specificity to improve the study of structure and function of laccase. Methods: According to the principle of homologous cloning, laccase gene from Arabidopsis thaliana was used to blast the database JGI of Populus trichocarpa . The Populus tri-chocarpa laccase PtrLAC -2 gene was cloned by PCR before being ligated with pET-30a(+) to con-struct prokaryotic expression vector PtrLAC-2-pET30a(+). PtrLAC-2-pET30a(+) was then trans-formed into E. coli BL21(DE3) competent cells for induction expression. Recombinant protein was purified as antigen to immune rabbit to prepare polyclonal antibody. Titer of the polyclonal anti-body and specificity were analyzed using ELISA and Western bolt at last. Results: Populus tricho-carpa laccase gene was isolated (renamed PtrLAC -2, Genebank: XP_002308164). Prokaryotic ex-pression vector PtrLAC-2-pET30a(+) was constructed successfully. The recombinant protein with the length of 62 KDa was obtained. ELISA analysis showed that the titer of the obtained antibody was 1:102,400. Western blot showed that the antibody could specifically combine with PtrLAC-2 protein in Populus trichocarpa . Conclusion: The PtrLAC -2 genes successfully expressed in E. coli and the PtrLAC-2 polyclonal antibody with high affinity and specificity was generated. This study will supply theoretical foundation in the following enzyme biochemical function. Keywords
Populus trichocarpa , Laccase, Gene Clone, Prokaryotic Expression, Polyclonal Antibody
毛果杨漆酶基因 PtrLAC -2的原核表达与多克隆 抗体制备研究
李 盈,李嘉鑫,滕晓瞳,郭晓蕊,程子义,尹 玢,张永卓,陆 海,李 慧

推荐阅读
- 超牛的招商银行营销话术
- 六年级健康教育试卷答案
- 2015年财政部财科所会计学考研复试重点内容,出题趋势分析
- Chapter 2 Care for hair(知识要点)课件(牛津深圳版九年级全册)
- 教师奖励基金实施办法建议书
- 提升职场竞争力经典实用课件之四十八:管理者的职业修炼
- 应急预案体系建设情况调查报告
- 我国港口城市的功能模式研究_陈航
- 公共关系 6套
- 案场礼仪规范与实操2
- hosts弹出广告屏蔽
- protel99se经典高校教程九
- 椰汁水晶冻香橙糕的做法
- 北京市文化创意产业战略研究与设计
- 第六章:力和机械测试题
- My,pen pal
- 西北工业大学电路基础课件第3章
- 《卓越领导力与执行力》
- ThinkPad BIOS设置中文手册(史上最全)
- 修改我的文档位置
- 最新人教版新目标九年级上期中考试英语试题及答案
- 儿童涂色画大全
- 将 Microsoft LoopBack Adapter 配置为系统的主网络适配器
- 严冰教授
- 文件签收单格式
- 2015年全国考纲词汇和短语--有音标
- 美国音乐种类英语ppt