Lee S G, Ricca G A, Crumley G, Lloyd R S, Drohan W
Meloy Laboratories, Inc., Rorer Biotechnology, Inc., Springfield, Virginia 22151.
Biochem Biophys Res Commun. 1988 Feb 29;151(1):598-607. doi: 10.1016/0006-291x(88)90636-5.
Plasmids expressing 2 forms of human immune interferon (IFN-gamma) in E. coli have been constructed: 1) pIFNTacI which expresses IFN-gamma with an N-terminal amino acid sequence of met-cys-tyr-cys-gln-, and 2) pIFNTacII which is a derivative of pIFNTacI from which the 9 base pairs (bp) coding for the cys-tyr-cys have been deleted. Quantitation of Western blots showed that approximately 10-fold more IFN-gamma was produced in cells harboring pIFNTacII (7.5% of total cellular protein) as compared to pIFNTacI. The IFN-gamma expressed in E. coli pIFNTacII is biologically active and routinely recoverable at 10(9) units per liter. When examined microscopically, IPTG induced E. coli harboring either plasmid construction contains prominent cytoplasmic inclusion bodies.
已构建出在大肠杆菌中表达两种形式人免疫干扰素（IFN-γ）的质粒：1）pIFNTacI，其表达的IFN-γ N端氨基酸序列为met-cys-tyr-cys-gln-；2）pIFNTacII，它是pIFNTacI的衍生物，编码cys-tyr-cys的9个碱基对（bp）已被删除。蛋白质印迹定量分析表明，与pIFNTacI相比，携带pIFNTacII的细胞中产生的IFN-γ约多10倍（占细胞总蛋白的7.5%）。在大肠杆菌pIFNTacII中表达的IFN-γ具有生物活性，通常每升可回收10⁹单位。在显微镜下检查时，IPTG诱导的携带任一质粒构建体的大肠杆菌都含有明显的细胞质包涵体。
