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以超速离心法提取的HepG2细胞总RNA为模板,应用RT-PCR技术扩增得到编码人干细胞因子(SCF)全长CDNA(约0.8kb)。克隆、序列分析确证编码人SCF膜外活性区的CDNA(约0.5kb)结构正确,构建了含有该基因的表达质粒(PBV-mhSCF)并在大肠杆菌中获得高效表达。
The total RNA of HepG2 cells extracted by ultracentrifugation was used as a template. The full-length CDNA encoding human stem cell factor (SCF) was amplified by RT-PCR (about 0.8 kb). Cloning and sequence analysis confirmed that the CDNA (about 0.5 kb) encoding the active region of human SCF was correctly constructed. The expression plasmid (PBV-mhSCF) containing this gene was constructed and highly expressed in E. coli.