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重组斑马鱼Gli2蛋白N端肽段的表达及纯化
谭训刚,张培军,杜少军
1.中国科学院海洋研究所;2.中国科学院研究生院
摘要:
Hedgehog(Hh)信号及其信号传递系统在生物机体发育中起着非常重要的作用。从Hhs分泌到其靶基因收到来自Hhs的信号的过程中,包括很多步骤,而每一步骤都有不同的因子参与作用。在其传递的过程中,参与受体细胞将胞外的信号传到胞内调节其目标基因的一个重要因子是果蝇分节-极性基因cubitus in terruptus(Ci)及其脊椎动物同源基 因-G li编码的蛋白。
关键词:  斑马鱼  重组蛋白  表达纯化
DOI:
分类号:
基金项目:国家自然科学基金项目(30210403202)资助
Expression and Purification of N-Terminal Partial Protein of Recombination Zebrafish Gli2
TAN Xungang1,2, ZHANG Peijun1, DU Shaojun3
1.Institute of Oceanology, The Chinese Academy of Sciences;2.Graduate School of the Chinese Academy of Sciences;3.Center of Marine Biotechnology, University of Maryland Biotechnology Institute
Abstract:
Based on the protein comparison between zebrafish G li2 and fruit fly Ci and the analysis of hydrophilicity/hydrophobicity of G li2 protein by computer programmer, the recombination N-terminal partial protein of G li2 protein was designed. After the DNA fragment encoding the N terminal part was cloned out by PCR, it was ligated with an expression vector containing H is-tag and transformed into BL-21, a bacteria l expression strain, to express this protein. After the expression condition was optimized, the protein can be expressed at high level after having been induced for 3 hours when the inducer— IPTG was at 0.6mmol /L. The protein is soluble in solution, a bacterial protein extraction solution. After the protein solution was diluted in B-per solution, the recombination protein can bind to the Ni-NTA column and be purified highly.
Key words:  N-Terminal  recombination protein  expression and purification
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