欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
目的：表达纯化胸腺嘧啶糖苷酶（TDG）蛋白并制备TDG多克隆抗体。方法：PCR扩增小鼠TDG（mTDG）的基因片段并插入pET28a（＋）原核表达载体，表达并纯化His—mTDG融合蛋白。用纯化的His-mTDG蛋白免疫兔子产生抗血清，进一步亲和纯化抗血清制备抗TDG的多克隆抗体，并检测分析制备的抗体在Westernblotting和免疫荧光分析中的应用。结果：在低温（20℃）和低IPTG浓度（0．2mmol·L^-1）时，His-mTDG融合蛋白大部分在可溶上清部分，用亲和层析法分离纯化了His—mTDG蛋白，并用纯化的蛋白制备了兔抗TDG抗体，结果显示制备的抗体能够特异性地在免疫分析中使用。结论：本研究纯化了条带单一的并且具有生物学活性的TDG融合蛋白，制备了具有良好特异性的兔抗TDG抗体，为进一步研究TDG的性质、结构和功能奠定了基础。
英文摘要：
Objective： To express and purify TDG protein, and to prepare the rabbit antibody against thymine DNA glycosylase（TDG） protein. Methods： Mouse TDC（mTDG） was amplified by PCR and then was inserted into the fusion expression vector pET28a（ ＋ ）. After being expressed in E. coli BL21, the His-mTDG protein was purified and used to immunize rabbit. Purified antibody was obtained through affinity chromatography column, and its applications were evaluated through Western blotting and immunofluorescence analysis. Results： His-mTDG fusion protein was almost all soluble at low temperature （ 20 ℃ ） and low IPTG condition （0.2 mmol· L^- 1 ）. The fusion protein was purified and used to immunize rabbits, the antibody against TDG was obtained. The further results showed that the antibody had a good specificity in immunoassays. Conclusion： We have purified mTDG fusion protein with an obviously homogeneous band and high biology activity and prepared the rabbit antibody against TDG successfully, which lays the foundation for further study on its character, structure and function.
同期刊论文项目
同项目期刊论文
A Contradictory Role of A(1) Adenosine Receptor in Carbon Tetrachloride- and Bile Duct Ligation-Indu
The Environmental Light Influences the Circulatory Levels of Retinoic Acid and Associates with Hepat
A novel genotyping strategy based on allele-specific inverse PCR for rapid and reliable identificati
Expression of Recombinant Human FADD, Preparation of Its Polyclonal Antiserum and the Application in
Impairment of Na(+), K(+) -ATPase in CD95 (APO-1) induced human T-cell leukemia cell apoptosis media
Enhancement of the antitumor properties of TRAIL by its targeted delivery to the tumor neovasculatur
A hybrid protein comprising ATF domain of pro-UK and VAS, an angiogenesis inhibitor, is a potent can
Fusion protein between protein ZZ and red fluorescent protein DsRed and its application to immunoass
Polyethylenimine-complexed plasmid particles targeting Focal Adhesion Kinase Function as Melanoma Tu
A novel ferritin gene, SferH-5, reveals heterogeneity of the 26.5-kDa subunit of soybean (Glycine ma
Cross-linked Polyethylenimine as Potential DNA Vector for Gene Delivery with High Efficiency and Low
AUF1-like protein binds specifically to DAS cis-acting element that regulates mouse alpha-fetoprotei
Combination of human plasminogen kringle 5 with ionizing radiation significantly enhances the effica
Fusion Protein between Protein ZZ and Enhanced Green Fluorescent Protein and its Application to Immu
期刊信息
位置：