Overlap-extension PCR Based Site-directed Mutants of UGRP1 Gene Promoter

BACKGROUND AIM: To construct sited-directed mutants of human UGRP1 gene promoter. MATERIALS AND METHODS: Mutants were constructed by overlap extention method. The plasmid pGL3-UGRP1(-112G) was used as the template,site-directed mutagenesis was performed by overlap-extention PCR method at -112 in UGRP1 gene promoter,and the expression vector of mutant at -112 was then constructed. RESULTS: By DNA sequencing, human UGRP1 gene promoter was successfully changed from G to A at -112 bp and the expression vector with the site-directed mutants were constructed. CONCLUSION: Overlap-extension PCR method was convenient and efficient for site-directed mutagenesis. Contruction of desired mutant pGL3-UGRP1(-112A) may provide a basis for the research on regulating transcriptional activity of G/A polymorphism at -112 bp of human UGRP1 gene promoter.