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. 1983 Apr;154(1):356–365. doi: 10.1128/jb.154.1.356-365.1983

In vitro construction and characterization of phoA-lacZ gene fusions in Escherichia coli.

S Michaelis, L Guarente, J Beckwith
PMCID: PMC217467  PMID: 6403507

Abstract

Using recombinant DNA techniques, we have constructed phoA-lacZ gene fusions. Two of the fusions encode hybrid proteins containing approximately half of alkaline phosphatase at the amino terminus joined to beta-galactosidase. For the one fusion strain analyzed in detail, it was shown that the hybrid protein is found in the membrane fraction of cells. In its membrane location, the beta-galactosidase activity of the hybrid is not sufficient to support cell growth on lactose. Unexpectedly, fusions containing phoA and lacZ joined in the wrong translational reading frame were also obtained. These fusions direct the phosphate-regulated synthesis of beta-galactosidase, apparently via a translation restart mechanism. Thus, when gene fusions are constructed, the presence of properly regulated beta-galactosidase activity does not necessarily indicate that a hybrid protein is being produced.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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