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人β-NGF基因在CHO细胞中表达的生物学活性及分离纯化 被引量:6
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作者 白艳艳 杨吉成 +5 位作者 缪竞诚 盛伟华 韩晓枫 李丽娥 谢宇锋 周忆新 《中国免疫学杂志》 CAS CSCD 北大核心 2003年第5期343-346,共4页
目的 :检测人 β NGF基因转染CHO细胞后培养上清中所表达的 β NGF的生物学活性及分离纯化。 方法 :采用脂质体介导的真核细胞转染 ,运用形态学观察和MTT法进行检测做定性定量分析 ,蛋白观察用SDS PAGE电泳。结果 :转染 β NGF基因的CH... 目的 :检测人 β NGF基因转染CHO细胞后培养上清中所表达的 β NGF的生物学活性及分离纯化。 方法 :采用脂质体介导的真核细胞转染 ,运用形态学观察和MTT法进行检测做定性定量分析 ,蛋白观察用SDS PAGE电泳。结果 :转染 β NGF基因的CHO细胞可分泌能促进PC12细胞生长的活性 β NGF ,且可透过血脑屏障。 结论 :转染 β NGF基因的CHO细胞所分泌的 β NGF ,接近自然合成的蛋白 ,具有较高生物学活性 ,为NGF的生物制药奠定了实验基础。 展开更多
关键词 β-ngf基因 CHO细胞 表达 生物学活性 分离 纯化
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β-NGF在真核细胞蛋白表达系统中的表达及活性鉴定
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作者 李贤慧 王桂云 +1 位作者 王育强 郭新民 《牡丹江医学院学报》 2002年第1期7-10,共4页
目的 :得到较高表达 ,且表达产物具有良好生物学活性的神经生长因子。方法 :将神经胶质源 β -NGF基因克隆于DNA转移载体pBacPAK8中 ,获得重组DNA转移载体pBacPAK -NGF ,与线性化Bm -BacPAK6修饰病毒基因组DNA共转染Bm -N细胞 ,经过体... 目的 :得到较高表达 ,且表达产物具有良好生物学活性的神经生长因子。方法 :将神经胶质源 β -NGF基因克隆于DNA转移载体pBacPAK8中 ,获得重组DNA转移载体pBacPAK -NGF ,与线性化Bm -BacPAK6修饰病毒基因组DNA共转染Bm -N细胞 ,经过体内重组 ,筛选到重组病毒。用重组病毒感染家蚕幼虫 ,经SDS-PAGE检测及利用PC12细胞进行NGF生物活力测定。结果 :感染重组病毒BmNPV -NGF5d的家蚕血淋巴有一条与天然NGF单体分子易相近的蛋白组分 ,且有类似于小鼠 2 .5gNGF的活性。结论 :神经生长因子在家蚕幼虫中得到较高表达 。 展开更多
关键词 真核表达系统 神经生长因子 生物活性 β-ngf基因 神经胶质源 基因表达 基因活性 中枢神经系统疾病
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Study on the Gene Transcription Level of Hippocampus NGF in C_(57) Mouse Development
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作者 刘晓军 吴梧桐 +1 位作者 张林元 高向东 《Journal of Chinese Pharmaceutical Sciences》 CAS 2000年第2期108-111,共4页
β-Nerve growth factor (β NGF) mRNA levels in hippocampa from C57BL/6J mice of different ages were compared by semi-quantitative RT-PCR method, taking β-actin gene as an internal control. The levels of 6-and 12-mon... β-Nerve growth factor (β NGF) mRNA levels in hippocampa from C57BL/6J mice of different ages were compared by semi-quantitative RT-PCR method, taking β-actin gene as an internal control. The levels of 6-and 12-month-old C57 mice were much lower compared with that of the l-month-old mouse. However, there was no significant difference between these two groups. This result suggests that gene transcription level of hippocampus β NGF in a C57, mouse descends evidently as it grows up. Base T at Site 294 of β NGF cDNA was substituted by C in the C57, mouse, as revealed by DNA sequencing for the first time. Nevertheless, this polymorphism of nucleotide did not make any difference in amino acid composition. 展开更多
关键词 NGF β-actin RT-PCR Gene transcription C57 mouse Development
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Study on the Gene Transcription Level of Hippocampus NGF in C57 Mouse Development
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作者 刘晓军 吴梧桐 +1 位作者 张林元 高向东 《Journal of Chinese Pharmaceutical Sciences》 CAS 2000年第2期108-111,共页
β-Nerve growth factor (β NGF) mRNA levels in hippocampa from C57BL/6J mice of different ages were compared by semi-quantitative RT-PCR method, taking β-actin gene as an internal control. The levels of 6-and 12-mon... β-Nerve growth factor (β NGF) mRNA levels in hippocampa from C57BL/6J mice of different ages were compared by semi-quantitative RT-PCR method, taking β-actin gene as an internal control. The levels of 6-and 12-month-old C57 mice were much lower compared with that of the l-month-old mouse. However, there was no significant difference between these two groups. This result suggests that gene transcription level of hippocampus β NGF in a C57, mouse descends evidently as it grows up. Base T at Site 294 of β NGF cDNA was substituted by C in the C57, mouse, as revealed by DNA sequencing for the first time. Nevertheless, this polymorphism of nucleotide did not make any difference in amino acid composition. 展开更多
关键词 NGF β-actin RT-PCR Gene transcription C57 mouse Development
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