期刊名称:Proceedings of the National Academy of Sciences
印刷版ISSN:0027-8424
电子版ISSN:1091-6490
出版年度:1990
卷号:87
期号:14
页码:5469-5473
DOI:10.1073/pnas.87.14.5469
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要: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.