Key Laboratory of Biotechnology of Antibiotics of Ministry of Health, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, No.1 Tiantan Xili, Beijing 100050, China.
Biotechnol Lett. 2011 Feb;33(2):253-61. doi: 10.1007/s10529-010-0428-0. Epub 2010 Oct 8.
Streptomyces is an attractive host for heterologous protein secretion. To further optimize its expression capacity, better expression vectors will be helpful. Here, based on pSGL1, a high copy number plasmid present in Streptomyces globisporus C-1027, we constructed a series of novel E. coli-Streptomyces shuttle expression vectors pIMB2-4. These vectors, which are compatible with pIJ-derived vectors, contain the strong promoter ermE*p and signal sequence SP (MelC1) of the first ORF of melanin operon in S. antibiotics (pIMB2), SP (CagA) of C-1027 apoprotein in S. globisporus C-1027 (pIMB3 and pIMB4). Using these vectors, human interleukin-6 (IL-6) could successfully be expressed and secreted using S. lividans TK24 as host. Furthermore, replacement of a rare leucine codon TTA with CTG in SP (CagA) enhanced IL-6 production.
链霉菌是异源蛋白分泌的理想宿主。为了进一步优化其表达能力，更好的表达载体将是有帮助的。在这里，基于链霉菌Globisporus C-1027 中存在的高拷贝数质粒 pSGL1，我们构建了一系列新型大肠杆菌-链霉菌穿梭表达载体 pIMB2-4。这些载体与 pIJ 衍生载体兼容，包含强启动子 ermE*p 和黑色素操纵子第一个 ORF 的信号序列 SP（MelC1）（pIMB2），链霉菌 Globisporus C-1027 中 C-1027 脱辅基蛋白的 SP（CagA）（pIMB3 和 pIMB4）。使用这些载体，使用链霉菌 lividans TK24 作为宿主，成功表达和分泌了人白细胞介素-6（IL-6）。此外，在 SP（CagA）中替换稀有亮氨酸密码子 TTA 为 CTG 增强了 IL-6 的产生。
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