1887

Abstract

High-level, inducible expression of heterologous genes in the cyanobacterium sp. strain PCC 7942 was obtained using the promoter and repressor. The gene of sp. strain PCC 7937 encoding plastocyanin precursor protein and the gene encoding β-glucuronidase were initially placed under the control of the promoter and repressor by cloning into the pTrc99C expression vector and were introduced into the chromosomal platform for integration in (PIM) of the R2-PIM9 recipient strain. These pTrc99C-derived constructs often gave rise to transformants that did not contain a complete insert gene, probably because of gene conversion events. Selection of the desired R2-PIM9 transformants was vastly improved using the new pTrcIS vector that contains the gene encoding streptomycin resistance as an extra antibiotic resistance marker. The influence of IPTG concentration and induction time on gene expression with the system in was determined using β-glucuronidase as a reporter. The PCC 7937 gene in was expressed to a high level upon induction with IPTG as shown by RNA and immunoblot analysis. The general usability of pTrcIS as a cloning vector for inducible heterologous gene expression in was confirmed by the introduction of several more genes.

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1995-04-01
2021-04-14
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