摘要
将人β－珠蛋白基因（２．４ｋｂ），分别反向克隆入逆转录病毒载体Ｎ_２β和Ｎ_２Ａ，并在Ｎ_２Ａ３＇ＬＴＲ插入４１２ｂｐ的增强子ＨＳⅡ（简称Ｎ_２ＡβＥ０．４）。将此两种逆转录病毒重组质粒分别转入（－２和ＰＡ３１７细胞，经Ｇ４１８筛选后，用ＰＡ３１７细胞产生的病毒粒子感染小鼠红白血病细胞ＭＥＬ。结果显示，人β－珠蛋白基因可在ＭＥＬ内得到表达，来自人ＨＳⅡ（４１２ｂｐ）的增强子可以增强人β－珠蛋白基因ｍＲＮＡ的表达水平。
The present result showed that humanβ-globin gene has been integrated into amphotropic packaging cell line PA3l 7 in 5/13 clones of PA317βand 1/14 clone of PA317βE0.4.Retrovirus titers of amphotropic recombinant retrovirus ranged from 10￣3 to 10￣4CFU/ml.MEL cells were transfected by supernant of PA3 1 7βand PA31 7 βE0.4.Northern blot showed that the expression of human β-globin gene in MELβE0.4 was significantly higher than MELβin mRNA level,This result indicated that an enhancer which comes from HS Ⅱ of human LCR can increase hurman β-globin gene mRNA expression in transfected MEL.However,the retrovirus titers of PA317β and PA317βE0.4 in our experiment were low.When large fragment enhancer(0.4kb)was inserted into retrovirus vector,it could bring about the human β-globin gene deletion in amphotropic packing cells.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
1994年第6期420-423,共4页
Acta Academiae Medicinae Sinicae
基金
"863"项目
