Construction of MHC class Ⅱ transactivator recombinant adenovirus vector

VernacularTitle:MHC Ⅱ类转激活因子基因的重组腺病毒载体的构建
Author: Minmin ZHANG ; Hua YANG ; Jing JIN ; Hongyu WU ; Zhaoshen LI
Publication Type:Journal Article
Keywords: Pancreatic neoplasms; Genes,MHC class Ⅱ; Gene therapy
From: Chinese Journal of Pancreatology 2009;9(6):399-401
CountryChina
Language:Chinese
Abstract: Objective To construct a recombinant adenovirus vector containing the gene of major histocompatibility complex(MHC)class Ⅱ transactivator(C Ⅱ TA).Methods The restriction fragment of CIITA was inserted into pUC57 vector with EeoR Ⅰ and Xho Ⅰ.Then,recombinant plasmid pShutde-GFP-CMV CⅡTA was constructed with EcoR Ⅰ and Sal Ⅰ,and was confirmed by restriction enzyme digestion and sequeneing.After the treatment with Ⅰ-Ceu Ⅰ and Ⅰ-See Ⅰ,the fragment C Ⅱ TA from recombinant plasmid DShuttle-GFP-CMV.CⅡTA Was inserted into vector pAdxsi.And the pAdxsi-GFP-C Ⅱ TA wag packed into liposome,and was transfected to 293 cens.Results Recombinant plasmid pShuttle-GFP-CMV-C Ⅱ TA Was constructed successfully. After packed into vector pAdxsi, and transfected to 293 cells, significant virus Dlaques were observed,which showed the successful homologous recombination.The titer of the purified AdC Ⅱ TA was 2.0×10~(11) PFU/ml.Conclusions Recombinant adenovirus AdC Ⅱ TA containing gene of MHC class Ⅱ transactivator was established successfully.