Rüther U, Tripodi M, Cortese R, Wagner E F
European Molecular Biology Laboratory, Heidelberg, FRG.
Nucleic Acids Res. 1987 Sep 25;15(18):7519-29. doi: 10.1093/nar/15.18.7519.
Alpha 1-antitrypsin (alpha 1AT) present in large amounts in human serum and synthesized predominantly in hepatocytes, is the most abundant protease inhibitor and alpha 1AT mutant proteins are associated with hereditary disorders. To investigate the regulation of the normal human alpha 1AT gene, we have microinjected fertilized mouse eggs with a 17.5 kb DNA fragment containing the entire gene with 7 kb 5' and 0.3 kb 3' flanking sequences. We show that this DNA fragment contains all the information for efficient, accurate and tissue-specific expression. High serum concentration of the human protein was found in three independent transgenic mouse lines. The human alpha 1AT RNA is transcribed efficiently in liver, kidney and macrophages and we demonstrate that two different promoters are used for the expression in liver and macrophages of the transgenic mice.
α1抗胰蛋白酶（α1AT）大量存在于人体血清中，主要在肝细胞中合成，是最丰富的蛋白酶抑制剂，α1AT突变蛋白与遗传性疾病相关。为了研究正常人α1AT基因的调控，我们用一个17.5kb的DNA片段显微注射受精的小鼠卵，该片段包含整个基因以及7kb的5'侧翼序列和0.3kb的3'侧翼序列。我们表明，这个DNA片段包含了高效、准确和组织特异性表达的所有信息。在三个独立的转基因小鼠品系中发现了高血清浓度的人蛋白。人α1AT RNA在肝脏、肾脏和巨噬细胞中高效转录，我们证明在转基因小鼠的肝脏和巨噬细胞中使用了两种不同的启动子进行表达。
