Peng J, Zhang J, Xu Y, Liu D, Zhang H
Institute of Basic Medical Sciences, CAMS, Beijing.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1994 Dec;16(6):420-4.
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.
目前的结果表明，在PA317β的13个克隆中有5个以及PA317βE0.4的14个克隆中有1个，人β-珠蛋白基因已整合到双嗜性包装细胞系PA317中。双嗜性重组逆转录病毒的滴度范围为10³至10⁴CFU/ml。用PA317β和PA317βE0.4的上清液转染MEL细胞。Northern印迹显示，在mRNA水平上，MELβE0.4中人β-珠蛋白基因的表达明显高于MELβ。该结果表明，来自人LCR HS II的增强子可增加转染的MEL中人β-珠蛋白基因mRNA的表达。然而，在我们的实验中，PA317β和PA317βE0.4的逆转录病毒滴度较低。当将大片段增强子（0.4 kb）插入逆转录病毒载体时，它可导致双嗜性包装细胞中人β-珠蛋白基因的缺失。
