High-level expression of single-chain Fv-Fc fusion protein in serum and egg white of genetically manipulated chickens by using a retroviral vector
High-level expression of single-chain Fv-Fc fusion protein in serum and egg white of genetically manipulated chickens by using a retroviral vector
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DOI：
10.1128/jvi.79.17.10864-10874.2005
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发表时间：
2005-09-01
影响因子：
5.4
通讯作者：
Iijima, S
中科院分区：
文献类型：
--
作者：
Kamihira, M;Ono, K;Iijima, S
我们在这里报告使用逆转录病毒载体生产重组蛋白的转基因鸡的一代。结果发现，当将基于莫洛尼鼠白血病病毒的逆转录病毒载体注射到胚盘阶段的鸡胚中时，转基因表达被抑制。当将浓缩的病毒溶液注射到孵育50至60小时后发育中的胚胎的心脏中时，在整个胚胎中观察到转基因表达，包括性腺。为了实际生产，将编码与人免疫球蛋白G1的Fc区融合的抗朊病毒单链Fv（scFv-Fc）的表达盒的逆转录病毒载体注射到鸡胚中。稳定孵化的鸟在其血清和蛋中以高水平（类似于5.6mg/ml）产生scFv-Fc。我们从用这种方法产生的转基因鸡中获得了转基因后代。转基因稳定整合到转基因后代的染色体中。转基因后代也在血清和卵中表达scFv-Fc。
We report here the generation of transgenic chickens using a retroviral vector for the production of recombinant proteins. It was found that the transgene expression was suppressed when a Moloney murine leukemia virus-based retroviral vector was injected into chicken embryos at the blastodermal stage. When a concentrated viral solution was injected into the heart of developing embryos after 50 to 60 h of incubation, transgene expression was observed throughout the embryo, including the gonads. For practical production, a retroviral vector encoding an expression cassette of antiprion single-chain Fv fused with the Fc region of human immunoglobulin G1 (scFv-Fc) was injected into chicken embryos. The birds that hatched stably produced scFv-Fc in their serum and eggs at high levels (similar to 5.6 mg/ml). We obtained transgenic progeny from a transgenic chicken generated with this procedure. The transgene was stably integrated into the chromosomes of transgenic progeny. The transgenic progeny also expressed scFv-Fc in the serum and eggs.