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. 1990 Jul;87(14):5469–5473. doi: 10.1073/pnas.87.14.5469

Site-directed mutagenesis reveals a liver transcription factor essential for the albumin transcriptional enhancer.

K S Zaret 1, J K Liu 1, C M DiPersio 1
PMCID: PMC54346  PMID: 2371282

Abstract

A method based on the polymerase chain reaction is described for constructing a clustered set of basepair changes, deletions, or insertions at any site on a DNA fragment. Advantages of the procedure are that virtually every product has the desired sequence alteration and that only a single round of polymerase chain reaction is required. We used this method to demonstrate that the binding of a specific liver nuclear protein, which we call eH-TF, is essential for the function of the enhancer of the mouse albumin gene. The eH-TF binding activity is hepatocyte-specific; it binds to a functional region of the albumin promoter and is distinct from other albumin promoter factors, and part of the eH-TF binding sequence, TGTTTGC, occurs in functional regulatory sites of other liver-specific genes.

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