Kwon Mo Sun, Koo Bon Chul, Choi Bok Ruyl, Lee Hoon Taek, Kim Young Hye, Ryu Wang-Shik, Shim Hosup, Kim Jin-Hoi, Kim Nam-Hyung, Kim Teoan
Department of Physiology, Catholic University of Daegu School of Medicine, Daegu, Republic of Korea.
Biochem Biophys Res Commun. 2004 Jul 23;320(2):442-8. doi: 10.1016/j.bbrc.2004.05.197.
In this work we demonstrated the successful production of transgenic chickens expressing the enhanced green fluorescence protein (EGFP) gene. Replication-defective recombinant retroviruses produced from vesicular stomatitis virus G glycoprotein pseudotyped retrovirus vector system were injected beneath the blastoderm of non-incubated chicken embryos (stage X). From 129 injected eggs, 13 chicks hatched after 21 days of incubation. All hatched chicks were found to express vector-encoded EGFP gene, which was under the control of the Rous sarcoma virus promoter and boosted post-transcriptionally by woodchuck hepatitis virus post-transcriptional regulatory element sequence. Green fluorescent signals, indicative of the EGFP gene expression, were detected in various body parts, including head, limb, eye, toe, and several internal organs. Genomic incorporation of the transgene was also proven by Southern blot assay. Our results show the exceptional versatile effectiveness of the EGFP gene as a marker in the gene expression-related studies which therefore would be very helpful in establishing a useful transgenic chicken model system for studies on embryo development and for efficient production of transgenic chickens as bioreactors.
在这项工作中，我们展示了成功生产出表达增强型绿色荧光蛋白（EGFP）基因的转基因鸡。从水泡性口炎病毒G糖蛋白假型逆转录病毒载体系统产生的复制缺陷型重组逆转录病毒，被注射到未孵化的鸡胚（X期）胚盘下方。在129枚注射过的鸡蛋中，有13只小鸡在孵化21天后出壳。所有出壳的小鸡都被发现表达载体编码的EGFP基因，该基因受劳斯肉瘤病毒启动子控制，并通过土拨鼠肝炎病毒转录后调控元件序列在转录后得到增强。在包括头部、肢体、眼睛、脚趾和几个内部器官在内的各个身体部位都检测到了指示EGFP基因表达的绿色荧光信号。Southern印迹分析也证实了转基因在基因组中的整合。我们的结果表明，EGFP基因作为一种标记物在基因表达相关研究中具有非凡的通用有效性，因此对于建立一个用于胚胎发育研究和高效生产作为生物反应器的转基因鸡的有用转基因鸡模型系统将非常有帮助。
