Abstract
An expression vector plasmid, designated pMCSβgal, was constructed to contain a multiple cloning site for insertion of gene fragments with potential regulatory function. This plasmid was designed to test promoter activity in transgenic mice, since digestion with Pst I will release all vector sequences, which could inhibit transgene expression in vivo (Jaenisch, 1988). When this vector, containing the amelogenin promoter, was used to make transgenic mice, expression was tissue specific and developmentally regulated similar to the endogenous gene (Chen et al., 1994). In addition, the herpes simplex virus (HSV) thymidine kinase (TK) minimal promoter was inserted into pMCSβgal to produce pTKβgal, leaving an Sph I upstream site available for insertion of gene fragments with potential enhancer or silencer function, which can be assayed following transfection into cultured cells.
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