Renkawitz R, Beug H, Graf T, Matthias P, Grez M, Schütz G
Cell. 1982 Nov;31(1):167-76. doi: 10.1016/0092-8674(82)90416-0.
We transferred a chicken lysozyme gene recombinant by microinjection into primary cultures of chicken oviduct cells. The recombinant gene is a fusion between the lysozyme promoter, including 1.4 kb of upstream sequences, and the coding region of the gene for SV40 T antigen (plys-T). The expression of plys-T is stimulated by the steroid hormones progesterone and dexamethasone, but not by estradiol. The number of oviduct cells expressing coinjected or separately injected control genes is not increased by steroids. A deletion mutant lacking the lysozyme sequences between -161 and +15 does not express T antigen, indicating that transcription of plys-T starts within the lysozyme promoter region. By screening different cell types we found that microinjected plys-T is expressed in chicken oviduct cells but not in chicken macrophages or fibroblasts or in rat II fibroblasts.
我们通过显微注射将一个鸡溶菌酶基因重组体导入鸡输卵管细胞原代培养物中。该重组基因是溶菌酶启动子(包括1.4 kb的上游序列)与SV40 T抗原基因(plys-T)编码区的融合体。plys-T的表达受类固醇激素孕酮和地塞米松的刺激,但不受雌二醇的刺激。注射共注射或单独注射的对照基因的输卵管细胞数量不会因类固醇而增加。一个缺失-161至+15之间溶菌酶序列的缺失突变体不表达T抗原,这表明plys-T的转录起始于溶菌酶启动子区域内。通过筛选不同的细胞类型,我们发现显微注射的plys-T在鸡输卵管细胞中表达,但在鸡巨噬细胞、成纤维细胞或大鼠II型成纤维细胞中不表达。