Construction and identification of recombinant adenovirus vector containing CTLA4Ig gene
- VernacularTitle:CTLA4Ig重组腺病毒载体的构建与体外表达
- Author:
Xiaoti GUO
;
Yubin DENG
;
Caisheng LU
;
Shunon LI
- Publication Type:Journal Article
- Keywords:
T-lymphocytes,cytotoxic;
Immunoglobulins;
Adenoviral vector;
Gene therapy
- From:
Chinese Journal of Pathophysiology
1989;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct a recombinant adenovirus expression vector containing CTLA4Ig gene.METHODS: The CTLA4Ig gene derived from the plasmid PCDNA3.0/CTLA4Ig by using polymerase chain reaction (PCR) was inserted into the backward position of cytomegalovirus (CMV) immediate early promoter of the shuttle plasmid (pAdTrack-CMV). After being identified by endonuclease, PCR and sequencing, the recombinant shuttle plasmid pAdTrack-CTLA4Ig was co-transformed into E.coli. BJ5183 cells with the adeoviral backbone plasmid pAdEasyl-1 to obtain the homologous recombination. The adenovirus was generated in 293 cells. A series methods such as PCR and fluorescence microscope was employed to identify the generated recombinant adenovirus. RESULTS: Recombinant CTLA4Ig adenoviruses were constructed and the titer of virus was generally up to 1.65?10 12 phaque forming units per liter (PFU/L). CONCLUSION: Success in constructing recombinant pAdTrack-CTLA4Ig will be the base of the further research on its expression in the mammalian cells, and be potenially used in the prevention of transplant rejection and autoimmunity diseases.
