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Construction of Expression Vectors for the High Level Expression in Actinomycetes
Lamichhane, Tek Narayan,Lamichhane, Rajan,Lee, Hei Chan,Liou, Kwangkyoung,Sohng, Jae Kyung 한국공업화학회 2003 응용화학 Vol.7 No.1
Two expression vectors pIBR100 and pIBR20 were constructed for the high level expression in actinomycetes. The pathway specific gene (actII-ORF4) present in pIBR100 along with bi-directional Pact promoter helps to regulate the expression of different genes in actinomycetes. The act promoter, associated RBS, and His-tag were kept at the upstream of the multiple cloning sitcs. Similarly in pIBR20, a strong constitutive promoter ermE* was cloned along with suitable multiple cloning sites with His-tag. It is a bi-replicate origin vector, which can be used for the expression in both E. coli and actinomycetes. The use of the vector system provides the means for overexpression of several Streptomyces genes and provides a ground for the development of several bioactive materials.