[Retroviral-mediated transfer of human beta-globin gene and its enhancer into MEL cells].
The present result showed that human beta-globin gene has been integrated into amphotropic packaging cell line PA317 in 5/13 clones of PA317 beta and 1/14 clone of PA317 beta E0.4. Retrovirus titers of amphotropic recombinant retrovirus ranged from 10(3) to 10(4) CFU/ml. MEL cells were transfected by supernant of PA317 beta and PA317 beta E0.4. Northern blot showed that the expression of human beta-globin gene in MEL beta E0.4 was significantly higher than MEL beta in mRNA level. This result indicated that an enhancer which comes from HS II of human LCR can increase human beta-globin gene mRNA expression in transfected MEL. However, the retrovirus titers of PA317 beta and PA317 beta E0.4 in our experiment were low. When large fragment enhancer (0.4 kb) was inserted into retrovirus vector, it could bring about the human beta-globin gene deletion in amphotropic packing cells.
Peng
J
J
Institute of Basic Medical Sciences, CAMS, Beijing.
Zhang
J
J
Xu
Y
Y
Liu
D
D
Zhang
H
H
D000818:Animals; D004274:DNA, Recombinant; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007942:Leukemia, Experimental / Q000235:genetics* / Q000473:pathology; D051379:Mice; D010957:Plasmids; D012333:RNA, Messenger / Q000096:biosynthesis; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
D004274:DNA, Recombinant; D012333:RNA, Messenger; D005914:Globins