Construction and application of a mouse HPRT targeting vector with Cre recombinase recognition site lox66 / 第二军医大学学报
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-677410
Responsible library:
WPRO
ABSTRACT
Objective:
To study the construction and application of a vector with Cre recombinase recognition site lox66 for mouse HPRT gene targeting in embryonic stem(ES) cells.Methods:
Using the HPRT genomic DNA fragment and synthesized oligonucleotides, pSP HPRT lox66 Neo was designed and constructed as a replacement vector by common molecular cloning techniques. After the linearized pSP HPRT lox66 Neo DNA was electroporated into ES cells, and transfected cells were cultured in G418/6 TG drug selection medium. The recombination efficiency of this vector was tested.Results:
The main components of pSP HPRT lox66 Neo were a positive selection gene Neo, lox66, long and short homologous fragments of mouse HPRT gene and plasmid backbone. Twenty ES cell clones with HPRT gene inactivated were obtained.Conclusion:
An effective replacement vector with Cre recombinase recognition site lox66 is constructed and applied to HPRT gene targeting in ES cells.
Full text:
Available
Database:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Academic Journal of Second Military Medical University
Year:
1985
Document type:
Article