Five plasmids (pUC-PDC1, pUC-PGK1, pUC-ADH1, pUC-TPI1 and
pUC-HOG1) were first constructed. Primers 4PDC1-U and 4PDC1-D
were used to amplify the PDC1 promoter, and this PCR fragment
was digested by HindIII-EcoRI. The digested fragment was inserted
into the same restriction sites of the pUC18, and the resulting
plasmid was pUC-PDC1. The PGK1 promoter was amplified by
primers 4PGK1-U and 4PGK1-D and then digested by HindIII-PstI.
The digested fragment was inserted into the same restriction sites
of the pUC18, and the resulting plasmid was pUC-PGK1. Primers
4ADH1-U and 4ADH1-D were used to amplify the ADH1 promoter,
and this PCR fragment was digested by HindIII-EcoRI. The digested
fragment was inserted into the same restriction sites of the pUC18,
and the resulting plasmid was pUC-ADH1. The TPI1 promoter was
amplified by primers 4TPI1-U and 4TPI1-D and then digested by