Molecular cloning, sequence analysis of mouse integrin ?_4 gene and repairing of its mutation / 第三军医大学学报

ABSTRACT

Objective To clone and analyze the full-length cDNA of mouse integrin ?4, and repair the mutation sensible locei that caused the change of amino acids. Methods The cDNA of ?4 gene was amplified by RT-PCR using the total RNA extracted from mouse small intestine peyer’s patch. The PCR product was inserted into pMD19-T vector and then transformed into E. coli JM109. The positive recombinant clone was analyzed by restriction endonuclease and DNA sequencing. The mutation of ?4 cDNA that caused the change of amino acids was repaired. Results The cDNA of mouse ?4 had a length of 3 099 bp, and encoded a product of 1 032 amino acids. There were 12 bases pairs mutation of ?4 gene and the 6 base pairs causing the change of amino acids was repaired. Conclusion The cDNA of mouse ?4 is cloned successfully.