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Journal of Virology, September 2005, p. 10864-10874, Vol. 79, No. 17
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.17.10864-10874.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

High-Level Expression of Single-Chain Fv-Fc Fusion Protein in Serum and Egg White of Genetically Manipulated Chickens by Using a Retroviral Vector

Masamichi Kamihira,1* Ken-ichiro Ono,1 Kazuhisa Esaka,1 Ken-ichi Nishijima,1 Ryoko Kigaku,2 Hiroyuki Komatsu,2 Takashi Yamashita,3 Kenji Kyogoku,3 and Shinji Iijima1*

Department of Biotechnology, Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya,1 Innovation Plaza Tokai, Japan Science and Technology Agency, Minami-ku, Nagoya,2 KANEKA Corporation, Takasago, Hyogo, Japan3

Received 19 March 2005/ Accepted 24 May 2005

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 (~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.

* Corresponding author. Mailing address for Dr. Iijima: Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan. Phone: 81-52-789-4275. Fax: 81-52-789-3221. E-mail: iijima{at}nubio.nagoya-u.ac.jp.Present address for Dr. Kamihira: Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Fukuoka 812-8581, Japan. Phone and fax: 81-92-642-3526. E-mail: kamihira{at}chem-eng.kyushu-u.ac.jp.

Journal of Virology, September 2005, p. 10864-10874, Vol. 79, No. 17
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.17.10864-10874.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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