对链霉菌表达体系pIJ459/S. lividans TK54中表达外源基因hGM-CSF的可行性进行了探索,分析了影响外源基因表达的某些因素如:SD与ATG之间的距离,时间对表达的诱导等.为获得外源基因的高效表达探索了最适条件,然后利用MEL信号肽与目的基因片段融合,在链霉菌中获得了分泌型hGM-CSF表达产物,其中有1/3直接分泌到培养基中,其纯化产物经TF-1细胞培养及集落形成实验均证实其生物学活性达到5MU/L.
The high level secretive expression of hGM-CSF in streptomyces was obtained.The factors, which can influence the exogenous expression in prokaryoties, such as the distance between SD and ATG, and the features of expression induction in streptomyces,etc. were studied. Then the secreted hGMCSF protein in streptomyces using the signal peptide MEL were obtained. One third of the expression products were secreted. By the experiment of clony stimulating factor formation and cell culture of TF-1 (hGM-CSF dependent cell line),its biological activity determined is 5 MU/L.
刘岱岚,强伯勤,袁建刚,李元,梁植权.人GM-CSF在链霉菌中分泌表达的研究[J].生物化学与生物物理进展,1996,23(1):56-59
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