Beermann F, Ruppert S, Hummler E, Bosch F X, Müller G, Rüther U, Schütz G
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
EMBO J. 1990 Sep;9(9):2819-26. doi: 10.1002/j.1460-2075.1990.tb07470.x.
The c-locus of the mouse is thought to encode tyrosinase, the key enzyme for melanin synthesis in melanocytes of the skin and the eye. Recently, a mouse cDNA was isolated and shown to confer tyrosine activity on a cell line which expressed no specialized functions for melanin synthesis. To verify that the isolated tyrosinase gene is encoded at the genetically well characterized c-locus, a minigene was assembled from tyrosinase cDNA and tyrosinase genomic DNA and used for generation of transgenic mice. Following microinjection of this construct into fertilized eggs of an albino mouse strain, transgenic mice were obtained which showed pigmentation in skin and eyes. By in situ hybridization, we show expression of the transgene in melanocytes of the hairbulb and in the pigmented cell layers of the eye. We conclude that we have rescued the albino mutation (c/c) by introduction and expression of a functional tyrosinase gene.
小鼠的c基因座被认为编码酪氨酸酶，它是皮肤和眼睛黑素细胞中黑色素合成的关键酶。最近，一个小鼠cDNA被分离出来，并显示出能赋予一个对黑色素合成无特殊功能的细胞系酪氨酸活性。为了验证分离出的酪氨酸酶基因是在遗传特征明确的c基因座上编码的，一个由酪氨酸酶cDNA和酪氨酸酶基因组DNA组装而成的小基因被用于生成转基因小鼠。将该构建体显微注射到一个白化病小鼠品系的受精卵中后，获得了在皮肤和眼睛中出现色素沉着的转基因小鼠。通过原位杂交，我们显示转基因在毛球的黑素细胞和眼睛的色素细胞层中表达。我们得出结论，通过导入和表达功能性酪氨酸酶基因，我们挽救了白化病突变（c/c）。
