人bFGF卵清蛋白分泌表达载体的构建及分析 |
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引用本文: | 王海军,王艳芳,江莺,赵文杰,任艳,马吉胜. 人bFGF卵清蛋白分泌表达载体的构建及分析[J]. 黑龙江医药, 2010, 23(3): 328-331 |
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作者姓名: | 王海军 王艳芳 江莺 赵文杰 任艳 马吉胜 |
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作者单位: | 吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林,长春,130118 |
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基金项目: | 吉林农业大学博士启动基金 |
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摘 要: | 目的:构建人碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的卵清蛋白分泌表达载体,为建立鸡输卵管生物反应器奠定基础。方法:利用PCR手段,从鸡基因组DNA克隆了4.1k(-4050bp-+42bp)卵清蛋白(ovalbumin)上游调控序列,利用重叠PCR技术,将溶菌酶信号肽(lysozyme signal peptide)基因与人碱性成纤维细胞生长因子(bFGF)基因融合在一起。结果:经DNA测序和核酸电泳检测,OV启动子基因序列、pcsbFGF和pcOV-sbFGF电泳住点与预计结果一致。结论:人bFGF的卵清蛋白分泌表达载体构建初步完成,可进行进一步转染表达。
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关 键 词: | bFGF 卵清蛋白 分泌表达载体 生物反应器 |
Construction and Analysis of Ovalbumin Secreted Vector for Human bFGF |
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Affiliation: | Wang Haijun, et al (Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education, Jilin Agriculture University (Changchun 130118)) |
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Abstract: | Objective: To construct a ovalbumin secreted vector for human basic fibroblast growth factor (bFGF), that will be used in Chicken Oviduct Bioreactor. Methods: We obtain 4.1k (-4050bp - +42 bp) ovalbumin upstream regulatory sequence from chicken genomic DNA using PCR method, and joint the chicken lysozyme signal peptide gene and human hFGF gene together using overlap PCR technology. Results: By DNA sequencing and Nucleic Acid Electro- phoresis assay, the OV promoter, pcsbFGF and pcOV-sbFGF have the same results of their design sequence respectively. Conclusions: The ovalbumin secreted vector for human bFGF construction is completed preliminary, and next it will be used for the secretion of bFGF in Chicken Oviduct Bioreaetor. |
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Keywords: | bFGF |
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