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[Vector construction for embryonic stem cell gene targeting of site specific point mutation mouse coagulation factor
1Department of Medical Genetics, The Second Military Medical University, Shanghai 200433 P.R.China. LiJian-1996@Yahoo.com
Objective:
To construct the recombinant vectors for embryonic stem(ES) cell gene targeting which contain the mouse coagulation factor IX (F IX) gene modified by PCR site-directed mutagenesis.
Methods:
Three site specific point mutations were introduced into exon 8 of mouse F IX gene respectively. The replacement targeting vectors were constructed and transfected into ES cells. The drug-resistant cell clones were picked after drug selection.
Results:
The construction of targeting vectors was successful and several drug-resistant ES cell clones were gained.
Conclusion:
The site specific point mutation system can modify human gene in vitro more accurately. It is useful in the setting up of animal models.