Carcinogenesis, Teratogenesis & Mutagenesis ›› 2007, Vol. 19 ›› Issue (1): 8-010.doi: 10.3969/j.issn.1004-616x.2007.01.003

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

CHENG Feng1,2,SHENG Yan2， SHI Jing-yi2, CHEN yi2, XU Chao2, LIU Zhi2,LIANG Jun2, ZHU Zhong-yong1, SONG Huai-dong2   

  1. 1. PLA Center for Laboratory Medicine, Fuzhou General Hospital,Nanjin Military Command, Fuzhou 350025,Fujian China;2. State Key Lab for Medical Genomics,Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Jiaotong University, Shanghai 200025
  • Received:2006-06-30 Revised:2006-09-22 Online:2007-01-30 Published:2007-01-30

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Abstract: 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.

Key words: human UGRP1 gene, promoter, overlap extention PCR, site-directed mutagenesis

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