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Expression of the E. coli LacZ Gene in Chicken Embryos Using Replication Defective Retroviral Vectors Packaged With Vesicular Stomatitis Virus G Glycoprotein Envelopes |
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Teoan Kim, Young Man Lee, Hoon Taek Lee, Young Tae Heo, Heng-Cherl Yom, Mo Sun Kwon, Bon Chul Koo, Key Whang, Kwang Soo Roh |
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| Abstract |
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Despite the high potency of the retrovirus vector system in gene transfer, one of the main drawbacks of has been difficulty in preparing highly concentrated virus stock. Numerous efforts to boost the virus titer have ended in unsatisfactory results mainly due to fragile property of retrovirus envelope protein. In this study, to overcome this problem, we constructed our own retrovirus vector system producing vector viruses encapsulated with VSV-G (vesicular stomatitis virus G glycoprotein). Concentration process of the virus stock by ultracentrifuge did not sacrifice the virus infectivity, resulting in more than 108 to 109 CFU (colony forming unit) per ml on most of the target cell lines tested. Application of this high-titer retrovirus vector system was tested on chicken embryos. Injection of virus stock beneath the blastoderms of pre-incubated fertilized eggs resulted in chick embryos expressing E. coli LacZ gene with 100% efficiency. Therefore, our results suggest that it is possible to transfer the foreign gene into chicken embryo using our high-titer retrovirus vector. |
| Keywords:
Retrovirus Vector; Vesicular Stomatitis Virus G Glycoprotein; E. coli LacZ Gene; Green Fluorescent Protein; Transgenic Chicken Embryo |
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