The preparation of recombinant rainbow trout insulin like growth factorsⅠ(IGF Ⅰ) andⅡ(IGF Ⅱ) using the His6 fusion polypeptide technique and the mitogen ic activities of these compounds were investigated. Rainbow ...The preparation of recombinant rainbow trout insulin like growth factorsⅠ(IGF Ⅰ) andⅡ(IGF Ⅱ) using the His6 fusion polypeptide technique and the mitogen ic activities of these compounds were investigated. Rainbow trout IGF Ⅰand IGF ⅡB C A D domain cDNA were subcloned into expression vector pET 15b (Nova gen, USA) and expressed in host E. coli BL21 (DE3) as fusion polypeptides co ntai ning a stretch of 6 histidines at the N terminus (referred as His6 fusion polyp e ptide). The addition of IPTG (0 4 mmol/L) induced the expression of polypeptide s with a molecular weight of about 10 0 kDa. The expressed proteins were ammoniu m sulfate fractionally precipitated and further purified using a Ni 2+ His ·Bind R esin (Novagen, USA) affinity column. The reduced SDS PAGE showed that IGF Ⅰ w as of 80% purity and IGF Ⅱ was of 90% purity. The rainbow trout IGF Ⅰ His6 fusi on polypeptide showed dose dependent mitogenic effects on BALB/NIH3T3 cells in th e serum free medium culture, and rainbow trout IGF Ⅱ His6 fusion polypeptide al s o showed this kind of activity, but with lower potency. Taken together,these re sults indicated that the rainbow trout IGF Ⅰ and ⅡHis6 fusion polypeptides pr o duced in E. coli were biologically active, with IGF Ⅰ His6 fusion polypept ide of higher potency.展开更多
很多体外试验结果证明,可溶性葡萄球菌A蛋白(SpA)是人和动物T淋巴细胞和B淋巴细胞的促有丝分裂原和多克隆激活物。T细胞的两个亚群——辅助性T细胞(helper T cell,Th)和抑制性T细胞(Supperssor T coil,Ts)是免疫调节的中心。SpA对活体的...很多体外试验结果证明,可溶性葡萄球菌A蛋白(SpA)是人和动物T淋巴细胞和B淋巴细胞的促有丝分裂原和多克隆激活物。T细胞的两个亚群——辅助性T细胞(helper T cell,Th)和抑制性T细胞(Supperssor T coil,Ts)是免疫调节的中心。SpA对活体的T淋巴细胞及其亚群的影响的研究具有重要的理论和临床意义。现将我们观察的结果报告如下。展开更多
文摘The preparation of recombinant rainbow trout insulin like growth factorsⅠ(IGF Ⅰ) andⅡ(IGF Ⅱ) using the His6 fusion polypeptide technique and the mitogen ic activities of these compounds were investigated. Rainbow trout IGF Ⅰand IGF ⅡB C A D domain cDNA were subcloned into expression vector pET 15b (Nova gen, USA) and expressed in host E. coli BL21 (DE3) as fusion polypeptides co ntai ning a stretch of 6 histidines at the N terminus (referred as His6 fusion polyp e ptide). The addition of IPTG (0 4 mmol/L) induced the expression of polypeptide s with a molecular weight of about 10 0 kDa. The expressed proteins were ammoniu m sulfate fractionally precipitated and further purified using a Ni 2+ His ·Bind R esin (Novagen, USA) affinity column. The reduced SDS PAGE showed that IGF Ⅰ w as of 80% purity and IGF Ⅱ was of 90% purity. The rainbow trout IGF Ⅰ His6 fusi on polypeptide showed dose dependent mitogenic effects on BALB/NIH3T3 cells in th e serum free medium culture, and rainbow trout IGF Ⅱ His6 fusion polypeptide al s o showed this kind of activity, but with lower potency. Taken together,these re sults indicated that the rainbow trout IGF Ⅰ and ⅡHis6 fusion polypeptides pr o duced in E. coli were biologically active, with IGF Ⅰ His6 fusion polypept ide of higher potency.
文摘很多体外试验结果证明,可溶性葡萄球菌A蛋白(SpA)是人和动物T淋巴细胞和B淋巴细胞的促有丝分裂原和多克隆激活物。T细胞的两个亚群——辅助性T细胞(helper T cell,Th)和抑制性T细胞(Supperssor T coil,Ts)是免疫调节的中心。SpA对活体的T淋巴细胞及其亚群的影响的研究具有重要的理论和临床意义。现将我们观察的结果报告如下。
