小鼠造血干细胞因子的cDNA克隆化及在大肠杆菌表达

cDNA CLONING OF MURINE STEM CELL FACTOK AND ITS EX-PRESSION IN E.COLI

造血干细胞因子（ＳＣＦ）是一种多功能造血生长因子，与其它造血生长因子协同作用，刺激不同分化阶段的造血细胞增殖和分化。本文采用ＲＴ－ＰＣＲ技术从新生ＢＡＬＢ／Ｃ小鼠胸腺组织克隆了ＳＣＦ膜外功能区ｃＤＮＡ，进而在大肠杆菌表达出具有生物学活性的重组蛋白。

n this study,murine SCF cDNA encoding extra-cellular domain was isolated using reverse-transcription polymerase chainreaction(RT-PCR)from thymus derived from a new-bornBALB/C mouse.For the first time,it was demonstrated that SCF mRNA was expressed inmouse thymus.With the SCF cDNA in hand,the construct pKpL-nKL4 expressing non-fusionSCF in E.coli was made,which produce recombinant SCF molecules in the form of inclusionbodies,Active forms of proteins were obtained through the process of denaturation and renatu-ration of recombinant SCF,as indicated by the colony formation assay with mouse and humanbone marrow cells.