Construction and identification of the eukaryotic co-expression vector of human HLA-DRB 1*0901 and HLA-DRA with pVITRO2
WANG Dongmei
SONG Changxing
ZHANG Zhixin
Abstract：With an aim of evaluating protecting effects of HLA-DRB1 on xenograft, we performed this study to construct the eukaryotic co-expression vector of human HLA-DRB1*0901 and HLA-DRA with pVITRO2 and identify its expression in kidney cell of porcine (LLC-PK1).RT-PCR was used to amplify the HLA-DRBl*0901 and HLA-DRA cDNA from total RNA of leukocyte in peripheral blood.The HLA-DRA DNA fragment (digested with Bgl Ⅱ ) was inserted into the multiple cloning site 2 of vector pVITRO2; the HLA-DRB1*0901 DNA fragment (digested with BamH Ⅰ ), whose direction was previously identified, was inserted into the multiple cloning site 1 of vector pVITRO2-dra.Then, the co-expression vector pVITRO2-dra-drb was transfected into the LLC-PK1, while its expression was identified with Westem-blotting.The restriction endonuclease digestion and PCR suggested that the co-expression vector pVITRO2-dra-drh was constructed successfully.Western-blotting confirmed the protein HLA-DRB1.The results confirms that he co-expression vec-tor pVITRO2-dra-drb can express human HLA-DRBl*0901 in LLC-PK1.
Keywords：LLC-PK1HLA-DRB 1Co-expression vector
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 40-42 )