人干细胞生长因子cDNA克隆表达和纯化及其造血种属特异性.docVIP

人干细胞生长因子cDNA克隆表达和纯化及其造血种属特异性 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：人干细胞生长因子cDNA克隆表达和纯化及其造血种属特异性 1 37℃融合表达的SDS检测 6 28℃诱导表达融合蛋白的存在形式分析 6 文2：结核分枝杆菌furB基因的克隆表达和纯化医学 8 1材料和方法 9 参考文摘引言： 13 原创性声明（模板） 14 文章致谢（模板） 14 正文 人干细胞生长因子cDNA克隆表达和纯化及其造血种属特异性 文1：人干细胞生长因子cDNA克隆表达和纯化及其造血种属特异性 Cloning，Expression and Purification of Human Stem Cell Growth Factor cDNA and Its Species-specificity in Hematopoiesis Abstract Stem cell growth factor (SCGF) is an early-acting hematopoitic cytokine that has two isoforms including hSCGF with full length molecules and hSCGFβ，78 amino acids of which lost in the coerved calcium-dependent carbohydrate-recognition domain (CRD).It has been demotrated that hSCGFβ is strictly species-specific in regulating study was aimed to explore whether human SCGF can exert synergistic stimulatory effect on heterogenous murine CFU-GM ，hSCGF cDNA was amplified from human fetal liver cDNA library by using two-step hSCGF mature peptide coding sequence was subsequently placed at dowtream of glutathione S-traferase (GST) sequence in GST gene fusion expression vector. The results indicated that there existed an additional 60 kD protein compared with mock BL21 when the cells hosting recombinant plasmid were induced with IPTG at 37℃.SDSanalysis demotrated that the GST-hSCGF fusion protein mainly existed in ioluble induced at low temperature (28℃)，the recombinant protein was mostly GST-fusion recombinant protein was subsequently purified by using affinity clonogenic assay revealed that，unlike hSCGFβ，hSCGF had the granulocyte/ macrophage promoting activity (GPA) for murine bone marrow GM progenitor. It is concluded that，in contrast to human SCGFβ，the intact molecular hSCGF may have no species specificity，implying that CRD domain in human SCGFβ does not directly bind to corresponding SCGF receptor，but may have certain biological function. Key words human stem cell growth factor; CRD region; cDNA clone; fusion expression; hematopoietic species speci