Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
J Biosci Bioeng. 2021 Apr;131(4):453-459. doi: 10.1016/j.jbiosc.2020.11.011. Epub 2020 Dec 24.
The use of promoters that strongly express target genes in the chicken oviduct is beneficial for the production of proteinaceous materials into egg white by transgenic chickens. To examine the regulatory mechanisms of chicken lysozyme gene expression in vivo, genetically manipulated chickens that express human erythropoietin under the control of a lysozyme promoter-enhancer were established. By using several deletion mutants of the promoter-flanking region, we found that a -1.9 kb DNase I hypersensitive site (DHS) was essential for oviduct-specific expression in genetically manipulated chickens. The concentration of human erythropoietin in egg white was 14-75 μg/ml, suggesting that the chicken lysozyme promoter containing -1.9 kb DHS is sufficient for the production of pharmaceuticals using transgenic chickens.
在转基因鸡中使用能够在鸡输卵管中强烈表达靶基因的启动子有利于将蛋白质材料生产到蛋清中。为了研究鸡溶菌酶基因在体内的表达调控机制,我们构建了在溶菌酶启动子增强子控制下表达人红细胞生成素的基因改造鸡。通过使用启动子侧翼区域的几个缺失突变体,我们发现-1.9kb 的 DNase I 超敏位点(DHS)对于基因改造鸡的输卵管特异性表达是必需的。人红细胞生成素在蛋清中的浓度为 14-75μg/ml,这表明含有-1.9kb DHS 的鸡溶菌酶启动子足以用于使用转基因鸡生产药物。