| 人角质细胞生长因子2真核表达载体的构建及功能验证 |
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| 引用本文： | 刘娟,赵思源,徐小洁,杨超,张鲁囡,刘佳,孙万军.人角质细胞生长因子2真核表达载体的构建及功能验证[J].生物技术通讯,2019(1):41-46. |
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| 作者姓名： | 刘娟 赵思源 徐小洁 杨超 张鲁囡 刘佳 孙万军 |
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| 作者单位： | 火箭军特色医学中心血液科;海军92076部队卫生队;军事医学研究院生物工程研究所;北京卫戍区第三退休干部休养所;解放军总医院第六医学中心感染控制科 |
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| 基金项目： | 国家自然科学基金(81472589;81672602;31600681);全军医学科技青年培育计划孵化项目(16QNP029);全军医学科技青年培育计划培育项目(17QNP029) |
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| 摘 要： | 目的:构建人角质细胞生长因子2(KGF-2)基因真核表达载体,探索KGF-2的上皮细胞迁移功能。方法:以人乳腺cDNA文库为模板,PCR扩增KGF-2基因片段,将其插入pXJ-40-myc,经双酶切和测序验证后,将重组质粒转染HEK-293T细胞,采用Western印迹检测重组蛋白;转染人肠上皮细胞FHC,采用细胞划痕实验检测细胞迁移能力。结果:PCR扩增获得627 bp的DNA产物,插入pXJ-40-myc载体,双酶切及测序结果证明重组质粒含有目的序列;转染HEK-293T细胞,Western印迹检测到相对分子质量约23×103的目的蛋白;细胞划痕实验显示,转染Myc-KGF-2的FHC细胞较未转染或转染空载体的细胞迁移能力强。结论:构建了人KGF-2基因真核表达载体,并验证了其促进细胞迁移的功能。
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| 关 键 词： | 人角质细胞生长因子2基因 真核表达 细胞迁移 |
Construction and Functional Verification of Human Keratinocyte Growth Factor-2 Eukaryotic Expression Vector |
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| LIU Juan,ZHAO Si-Yuan,XU Xiao-Jie,YANG Chao,ZHANG Lu-Nan,LIU Jia,SUN Wan-Jun.Construction and Functional Verification of Human Keratinocyte Growth Factor-2 Eukaryotic Expression Vector[J].Letters in Biotechnology,2019(1):41-46. |
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| Authors: | LIU Juan ZHAO Si-Yuan XU Xiao-Jie YANG Chao ZHANG Lu-Nan LIU Jia SUN Wan-Jun |
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| Institution: | (Department of Hematology,Rocket Military Characteristic Medical Center,Beijing 100088;Navy 92076 ArmyHealth Team,Beijing 102202;Beijing Institute of Biotechnology,Beijing 100850;Third Retired Cadres'Sana.torium of Beijing Garrison District,Beijing 100039;Department of Infectious Diseases,Six Medical Center ofPLA General Hospital,Beijing 100048,China) |
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| Abstract: | LIU Juan;ZHAO Si-Yuan;XU Xiao-Jie;YANG Chao;ZHANG Lu-Nan;LIU Jia;SUN Wan-Jun(Department of Hematology,Rocket Military Characteristic Medical Center,Beijing 100088;Navy 92076 ArmyHealth Team,Beijing 102202;Beijing Institute of Biotechnology,Beijing 100850;Third Retired Cadres'Sana.torium of Beijing Garrison District,Beijing 100039;Department of Infectious Diseases,Six Medical Center ofPLA General Hospital,Beijing 100048,China) |
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| Keywords: | human keratinocyte growth factor-2(KGF-2)gene eukaryotic expression cell migration |
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