关键词: &beta, 地中海贫血；CD41-42突变基因；重组载体；HeLa细胞

Abstract:

Abstract:Objective
To construct the  recombinant vector pEGFP-C2-CD41-42 of  CD41-42 mutation β thalassemia and  HeLa cell model,and  to provide  basis for cell and mouse model establishment of  CD41-42 mutation β thalassemia and the further gene therapy.Methods The DNA in peripheral blood of  βCD41-42 homozygote mutation patient was extracted and the DNA framen of β-globin  containing β CD41-42 was amplified by PCR,then the PCR product β CD41-42 globin gene  was subcloned into pEGFP-C2 vector.The recombinant plasmid pEGFP-C2-βCD41-42 was  transfected into HeLa cells by LipofectamineTM 2000.The fluorescence expression of βCD41-42 in HeLa cells after transfecton was observed.The expression of βCD41-42 in HeLa cells was identified by RT-PCR method.Results The   HeLa cells expressed green fluorescence  24 h after transfected with the recombinant plasmid  pEGFP-C2-βCD41-42   and expressed lots of green fluorescence after G418 screening.The special PCR products(227 bp)  was obtained by RT-PCR,but it didn’t express  in the HeLa cells transfected with pEGFP-C2 blank plasmid.Conclusion The recombinant vector  pEGFP-C2-βCD41-42  of CD41-42 mutation β thalassemia is successfully estabished and the HeLaCD41-42 cell model  is successfully established.

Key words: &beta, thalassemia；CD41-42 mutation gene；recombinant vector；HeLa cells