Townes T M, Lingrel J B, Chen H Y, Brinster R L, Palmiter R D
EMBO J. 1985 Jul;4(7):1715-23. doi: 10.1002/j.1460-2075.1985.tb03841.x.
Transgenic mice carrying human beta-globin genes were produced by microinjecting linear DNA molecules containing cloned beta-globin genes with up to 4300 bp of 5'-flanking sequence and 1700 bp of 3'-flanking sequence. Most (15 of 20) of these transgenic mice expressed the human beta-globin genes in blood cells and the level of expression in some mice was comparable with that obtained from endogenous beta-globin genes. Human beta-globin gene expression appeared to be restricted to cells of the erythroid lineage and was first detected between 11 and 14 days of development, in parallel with mouse beta-globin. Constructs with as little as 48 bp of 5'-flanking sequence also appeared to be expressed appropriately. The mRNA transcripts had correct 5' ends and directed human beta-globin synthesis in reticulocyte lysates. Human beta-globin protein was detectable in mature erythrocytes from progeny of one of these mice. The frequency and extent of expression was severely depressed when the procaryotic vector DNA was not removed prior to microinjection.
通过显微注射线性DNA分子来制备携带人类β-珠蛋白基因的转基因小鼠，这些线性DNA分子包含克隆的β-珠蛋白基因，其具有长达4300 bp的5'侧翼序列和1700 bp的3'侧翼序列。这些转基因小鼠中的大多数（20只中的15只）在血细胞中表达人类β-珠蛋白基因，并且一些小鼠中的表达水平与从内源性β-珠蛋白基因获得的水平相当。人类β-珠蛋白基因的表达似乎仅限于红系谱系的细胞，并且在发育的11至14天之间首次被检测到，与小鼠β-珠蛋白同时出现。含有低至48 bp的5'侧翼序列的构建体似乎也能正确表达。mRNA转录本具有正确的5'末端，并在网织红细胞裂解物中指导人类β-珠蛋白的合成。在其中一只小鼠后代的成熟红细胞中可检测到人类β-珠蛋白。当在显微注射前未去除原核载体DNA时，表达的频率和程度会严重降低。
