Construction of AT2R gene recombinant adenovirus by an efficent method / 医学研究生学报
Journal of Medical Postgraduates
;
(12)2003.
Article
in Chinese
| WPRIM
| ID: wpr-584791
ABSTRACT
Objective:
To construct the recombinant adenovirus of AT2R by using the method of homogenous recombination in bacteria.Methods:
AT2R gene was get from the vector of PUHD-AT2R by PCR ,and subcloned into shuttle vector of pAdTrack-CMV, forming transfer vector of pAdTrack-CMV/AT2R.Then it was linearized with PmeⅠ and transformed into Adeasier-1 cell. The DNA of identified recombinant plasmid was digested with PacⅠ and transfected to 293cells to package adenovirus. The PCR technique was used to detect target gene . The titre of the Ad-AT2R was measured with the aid of GFP expression.Results:
PCR test indicated each the recombinant adenovirus contained the insert of AT2R. The titre of purified recombinant adenovirus was 1.5?10 12pfu/ml.Conclusion:
The method of homologous recombination in bacteria is more convenient and efficient compared with that in cell .The prepared Ad-AT2R paves a sound foundation for further study.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Journal of Medical Postgraduates
Year:
2003
Type:
Article
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