Construction and expression of recombinant adenovirus vector encoding human somatostatin receptor type 2
- VernacularTitle:携带人生长抑素2型受体的腺病毒载体的构建和表达
- Author:
Bin XU
;
Hao XU
;
Yueqin LI
;
Zhongjian CAO
;
Xin ZHANG
;
Tianhong ZHOU
- Publication Type:Journal Article
- Keywords:
Receptors,somatostatin;
Adenoviridae;
Gene therapy;
Carcinoma
- From:
Chinese Journal of Pathophysiology
2000;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct recombinant adenovirus vector carrying the gene of human somatostatin receptor type 2(SSTR2) for gene therapy of pancreatic carcinoma.METHODS: SSTR2 cDNA was inserted into adenovirus shuttle plasmid pDC316,named pDC316-SSTR2.pDC316-SSTR2 was cotransfected with rescue plasmid pBHGlox(delta) E1,3Cre into 293 cells by liposome reagent.Ad-SSTR2 was generated by site-specific recombination and confirmed by PCR.Ad-SSTR2 was propagated in 293 cells and purified.The titer of viral stock was determined by end-point dilution assay.Western blotting was used to determine the expression of SSTR2 protein after human pancreatic carcinoma cell capan-2 was infected with recombinant adenovirus.RESULTS: pDC316-SSTR2 was successfully constructed.Recombinant adenovirus Ad-SSTR2 was acquired by pDC316-SSTR2 and pBHGlox(delta) E1,3Cre cotransfected into 293 cells.Ad-SSTR2 was characterized by PCR.The virus titer was 6.0?10~(12) pfu/L.SSTR2 protein was detected after adenovirus infected capan-2 48 h with Western blotting.CONCLUSION: The recombinant adenovirus vector encoding human SSTR2 is successfully constructed and correctly expressed in pancreatic carcinoma cells.This investigation provides the basis for study of gene therapy of pancreatic carcinoma.
