Humphries R K, Ley T, Turner P, Moulton A D, Nienhuis A W
Cell. 1982 Aug;30(1):173-83. doi: 10.1016/0092-8674(82)90023-x.
We have compared the function of the human alpha-, beta- and delta-globin genes using various plasmid expression vectors derived from pBR322. Amplification of recombinants occurred after their introduction, by calcium-phosphate-mediated DNA transfer, into monkey kidney cells that constitutively produce T antigen (COS cells). The human alpha-globin gene promoter functioned independently, but the beta-globin gene promoter was nearly totally dependent on the enhancing activity of the 72 bp direct repeats from the SV40 genome. Furthermore, when the human alpha- and beta-globin genes were linked in the same vector, the alpha promoter was active but the beta promoter was not. Function of the delta-globin gene promoter also depended on the enhancer element. In vectors containing the 72 bp repeats and the beta- or delta-globin gene, the activity of the beta-globin gene was approximately 50 times greater than that of the delta-globin gene, approximating the ratio of beta and delta mRNA observed in normal human bone marrow cells.
我们使用源自pBR322的各种质粒表达载体,比较了人类α-、β-和δ-珠蛋白基因的功能。通过磷酸钙介导的DNA转移将重组体导入组成型产生T抗原的猴肾细胞(COS细胞)后,重组体发生了扩增。人类α-珠蛋白基因启动子独立发挥作用,但β-珠蛋白基因启动子几乎完全依赖于来自SV40基因组的72 bp直接重复序列的增强活性。此外,当人类α-和β-珠蛋白基因连接在同一载体中时,α启动子具有活性,但β启动子没有活性。δ-珠蛋白基因启动子的功能也依赖于增强子元件。在含有72 bp重复序列以及β-或δ-珠蛋白基因的载体中,β-珠蛋白基因的活性比δ-珠蛋白基因大约高50倍,接近正常人骨髓细胞中观察到的β和δ mRNA的比例。