摘要
干细胞因子（SCF）是一种重要的造血生长因子，它在自体造血干细胞动员、肿瘤放疗化疗的辅助治疗以及其它血液病的治疗上具有良好的应用前景。为了制备高纯度、活性的重组人SCF，将人工合成的SCF的cDNA序列，克隆入原核表达载体pET43．1a中，转化大肠埃希菌BL21（DE3），获得了高效表达菌株。SCF通过诱导表达形成包涵体，包涵体经洗涤、变性和初步纯化后，进行直接稀释复性，复性液经离子交换层析、分子筛层析等分离除去共价二聚体和异构体，获得纯度大于95％的重组人SCF样品，生物学比活性在7．0×10^5U／mg以上。结果表明：建立了有效的rhSCF制备工艺，且产物具有较高的纯度和生物学活性。
Stem cell factor （SCF） is an important hematopoietic growth factor, it has good prospects in the applicalions of autogenous hematopoiesis of stem cell mobilization, in auxiliary therapy of radiotherapy and chemotherapy of tumor, and in therapy of blood diseases. In order to prepare highly pure and active rhSCF, cDNA sequence of artificially synthesized SCF was cloned into prokaryotie expression vector pET43, la to transform E. eoli BL21 （DE3） and obtained a highly effective expression strain. An inclusion body express-formed through inducing SCF and after washed, denatured, and initially purified was renatured direetty through dilution. Covalent dime＇r and isomer in the renatured liquid was isolated and removed through ion exchange chromatography, molecular sieve chromatography and other means. And obtained rhSCF sample of purity above 95% with bio-speeific activity at over 7.0 × 105 U/mg.he results showed that the effective preparation technology of rhSCF had been established, and the product had fairly high purity and biological activity.
出处
《微生物学杂志》
CAS
CSCD
2009年第5期30-34,共5页
Journal of Microbiology
关键词
重组人干细胞因子
原核表达
纯化
共价二聚体
生物学比活性
recombinant human stem cell factor （rhSCF）
prokaryotie expression
purify
covalent dimer
bio-specific activity
