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一种用于蓝藻聚球藻PCC 6301的新型表达载体。

A novel expression vector for the cyanobacterium, Synechococcus PCC 6301.

作者信息

Takeshima Y, Sugiura M, Hagiwara H

机构信息

Hagiwara Institute of Health, Japan.

出版信息

DNA Res. 1994;1(4):181-9. doi: 10.1093/dnares/1.4.181.

Abstract

A cyanobacterial expression vector was constructed using ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) promoter and terminator sequences derived from Synechococcus PCC 6301. recombinant plasmid, designated pARUB19, has an ampicillin-resistant (ApR) gene as a selectable marker and four unique restriction sites to allow the insertion of foreign genes. Using this vector, the luciferase gene from the firefly, Photinus pyralis, was introduced into Synechococcus PCC 6301 cells. The luciferase expression vector could be maintained stably in the host cells. Light production of luciferin/luciferase was detected in the transformants. Luciferase amounted to 1.2% of the total soluble protein. This plasmid may facilitate higher levels of foreign gene expression in Synechococcus PCC 6301.

摘要

利用源自聚球藻PCC 6301的1,5-二磷酸核酮糖羧化酶/加氧酶(RuBisCO)启动子和终止子序列构建了一种蓝藻表达载体。重组质粒命名为pARUB19,具有氨苄青霉素抗性(ApR)基因作为选择标记和四个独特的限制性酶切位点,以允许插入外源基因。使用该载体,将来自萤火虫Photinus pyralis的荧光素酶基因导入聚球藻PCC 6301细胞。荧光素酶表达载体能够在宿主细胞中稳定维持。在转化体中检测到荧光素/荧光素酶的发光。荧光素酶占总可溶性蛋白的1.2%。该质粒可能有助于在聚球藻PCC 6301中实现更高水平的外源基因表达。

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