人干细胞因子基因大肠杆菌表达产物结构分析及活性研究  

Structure analysis and bioactivity study of human stem cell factor gene expressed in Escherichia coli

作　　者：吴洽庆[1] 刘忠荣 张思仲[1] 马用信[1] 雒蓬轶 王红霞[3] 陈松森[4] 

机构地区：[1]四川大学华西医院医学遗传研究室,四川成都610041 [2]成都地奥制药集团有限公司,四川成都610041 [3]国家生物医学分析中心,北京100850 [4]中国医学科学院基础医学研究所,北京100005

出　　处：《中国药学杂志》2003年第10期798-801,共4页Chinese Pharmaceutical Journal

基　　金：国家"十五"计划资助课题;"863"计划资助课题 (2 0 0 1AA2 15 40 1)

摘　　要：目的　确证人干细胞因子基因在大肠杆菌表达产物的正确性。方法　通过蛋白质杂交 ,氨基酸末端序列分析 ,质谱分析及动物体内生物活性的研究 ,对人干细胞因子基因大肠杆菌表达产物———重组人干细胞因子 (rhSCF)分子进行了鉴定。结果　大肠杆菌表达产物rhSCFN端 15个氨基酸序列及C端 2个氨基酸序列与理论推测一致 ,产物的相对分子质量为 185 83.75 ,质量肽谱与理论序列的重叠率为 98.2 % ,动物试验亦表明当与粒细胞集落刺激因子 (G CSF)联用具有显著的外周血干(祖 )细胞动员作用。结论　人干细胞因子基因大肠杆菌表达产物与理论推测完全一致 ,为其中试及临床试验奠定了基础。OBJECTIVE: To identify the product of recombinant human stem cell factor(rhSCF) gene expressed in Escherichia coli. METHODS: The purified product was characterized through Western blot, terminal amino acid sequence analysis, mass spectrometry and in vivo bioactivity assay. RESULTS: The molecular weight of rhSCF is 18 583. 75. The N-terminal 15 amino acids sequence, C-terminal 2 amino acids sequence and its peptide mass mapping were conformable to the rhSCF gene. The experiments in monkeys also indicated that the rhSCF used with G-CSF had remarkable ability to mobilize the hematopoietic progenitor cell into peripheral blood. CONCLUSION: The expression of the rhSCF gene in E. coli is just as expected, which set a solid base for its scale-up and clinical trail.

关 键 词：人干细胞因子 基因表达 大肠杆菌 鉴定 粒细胞集落刺激因子 

分 类 号：R378.21[医药卫生—病原生物学] R394[医药卫生—基础医学]