Carcinogenesis, Teratogenesis & Mutagenesis ›› 2003, Vol. 15 ›› Issue (1): 10-12.doi: 10.3969/j.issn.1004-616x.2003.01.003

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CONSTRUCTION OF THE EXPRESSION VECTOR WITH THE MUTANT APOCⅡ PROMOTER USING SITE- DIRECTED MUTAGENESIS

CHEN Chun- hua 1, CAO Ying- lin 1, HU Wei- cheng 2, ZHAO li 1   

  1. 1. Institute of Immunology; 2. Institute of Pathology and Pathophysiology, School of Medicine, Shandong University, Jinan 250012 , China
  • Received:2002-07-23 Revised:2002-09-28 Online:2003-01-30 Published:2003-01-30
  • Contact: CAO Ying- lin

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Abstract: Purpose: To construct the expression vector with the mutant apolipoprotein CⅡ(apoCⅡ) promoter for studying the transcript activity of the mutant apoCⅡ promoter on - 190 base pair. Methods: The plasmid pGL 3 with normal apoCⅡ promoter was used as the template, and a pair of completely complementary primers with the desired mutation was designed, which make the plasmid amplified and mutated on - 190 base pair of the apoCⅡ promoter using Stratagene's site- directed mutagenesis kit. After being transformed into XL- Blue supercompetent cells, the nicked mutant plasmid was repaired in these cells. Results: The normal apoCⅡ promoter was successfully changed from T to A on the - 190 base pair, the expression vector with the mutant apoCⅡ promoter was constructed. Conclusion: The whole circled plasmid with the desired mutation was obtained by a rapid and efficient method for site- directed mutagenesis, which has an implication for detecting the activity of the mutant apoCⅡ promoter.

Key words: Apolipoprotein CⅡ, Site- directed mutation, Promoter