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Ubiquitous GFP expression in transgenic chickens using a lentiviral vector.

机译:使用慢病毒载体在转基因鸡中无处不在的GFP表达。

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We report the first ubiquitous green fluorescent protein expression in chicks using a lentiviral vector approach, with eGFP under the control of the phosphoglycerol kinase promoter. Several demonstrations of germline transmission in chicks have been reported previously, using markers that produce tissue-specific, but not ubiquitous, expression. Using embryos sired by a heterozygous male, we demonstrate germline transmission in the embryonic tissue that expresses eGFP uniformly, and that can be used in tissue transplants and processed by in situ hybridization and immunocytochemistry. Transgenic tissue is identifiable by both fluorescence microscopy and immunolabeling, resulting in a permanent marker identifying transgenic cells following processing of the tissue. Stable integration of the transgene has allowed breeding of homozygous males and females that will be used to produce transgenic embryos in 100% of eggs laid upon reaching sexual maturity. These results demonstrate that a transgenic approach inthe chick model system is viable and useful even though a relatively long generation time is required. The transgenic chick model will benefit studies on embryonic development, as well as providing the pharmaceutical industry with an economical bioreactor.
机译:我们报告了使用慢病毒载体方法在雏鸡中第一个普遍存在的绿色荧光蛋白表达,其中eGFP在磷酸甘油激酶启动子的控制下。先前已报道了使用能产生组织特异性但并非普遍存在的标志物的小鸡种系传播的几种证明。使用杂合子雄性的胚胎,我们证明了胚系在胚胎组织中的传递,可以均匀地表达eGFP,并且可以用于组织移植,并可以通过原位杂交和免疫细胞化学进行加工。可通过荧光显微镜和免疫标记法鉴定转基因组织,从而在组织加工后产生永久性标记,可鉴定转基因细胞。转基因的稳定整合允许纯合的雄性和雌性育种,这些雌性将在达到性成熟时产下100%的卵中产生转基因胚胎。这些结果表明,即使需要相对长的产生时间,在小鸡模型系统中转基因方法也是可行和有用的。转基因小鸡模型将有益于胚胎发育的研究,并为制药业提供经济的生物反应器。

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