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1 46(3) 185~192, 綜論報告利用基因轉殖家禽產製功能性蛋白質 (1) (1)(2) (1) (2) sding@ntu.edu.tw

2 186 DNA Recombinant DNA DNA 18 Harvey et al., Harvey et al., 2002; Kim et al., 2005; Poynter et al., Stage X 40,000 60,000 Blastoderm Area pellucida Subgerminal cavity DNA 1 Stage X primordial germ cells, PGCs gastrulation PGCs primitive streak primitive genital ridge) Hamburger and Hamilton, 1951 PGCs PGCs PGCs ppgcs PGCs cpgcs PGCs gpgcs PGCs Savva et al PGCs cpgcs

3 187 donor recipient PGCs germline chimera PGCs Ono et al., 1996 gpgcs cpgcs gpgcs feeder cells gpgcs Shin et al., 2008; Motono et al., 2010 RNA RNA DNA DNA DNA DNA Lillico, 2005 Lentivirus gag pol env Park, 2007 F0 Founder Poynter et al., 2009 germline transmission Kwon et al., Table 1 Differences between two infection methods in generating transgenic poultry Method Category Lentivirus gpgcs DNA acceptor Subgerminal cavity cells primordial germ cells Reference DNA delivery Injecting the Lentivirus into the subgerminal cavity of eggs in stage X Injecting the transfected PGCs into the two-days embryo Technical threshold low high (Harvey et al., 2002) (Kim et al., 2005) Transgenesis efficiency (Shin et al., 2008) (%) (Kwon et al., 2010) - 10 (Motono et al., 2010)

4 Lillico et al., 2005 Albumen, egg white 60% 99% ovalbumin, OVA 60 54% 2 OVA OVA 2008 OVA promoter Lillico et al OVA 2.8 kb Estrogen response element ERE 426 μg/ml; Kwon et al kb OVA 234 ng/ml Parket al kb OVA ERE 60 ng/ml OVA OVA OVA promoter 2 Table 2 Composition of a chicken egg Constituent Proportion of egg by weight (%) Components of individual constituents (%) Albumin Yolk Shell 10 Major albumen proteins in a 60 g egg Constituent Weight (g) Fraction of total protein (%) Ovalbumin Ovotransferrin Ovomucoid Lysozyme (Lillico et al., 2005)

5 189 3 OVA promoter Table 3 Production ofexogenousproteins driven by OVA promoter in transgenic poultry Production Species OVA promoter Enhancer Target protein Reference (per ml egg white) White Leghorn 2.8 kb 675 bp ERE hifn 1a 426 μg (Lillico et al., 2008) Japanese quail 1.35 kb rhil1rn 234 ng (Kwon et al., 2010) White Leghorn 3.5 kb 647 bp ERE hegf 60 ng (Park et al., 2015) ERE: Estrogen Response Element; hifn 1a: human interferon beta 1; rhil1rn: recombinant human interleukin 1 receptor antagonist; hegf: human epidermal growth factor. infundibulum magnum Whittow, 1999 isthmus shell gland ciliated non-secretory cells non-ciliated secretory cells Jung et al., 2011 tubular gland cells Type A Type B Type C Type A cell Wyburn et al., 1970 secretory granule Type C cell Type A cell Type B cell Wyburn et al., 1970 Wyburn et al. 1970

6 190 DNA chorioallantoic membrane, CAM DNA PCR DNA DNA gel filtration SDS-PAGE Western Blotting Whittow, G Sturkie's Avian Physiology. p574. Hamburger, V., and H. L. Hamilton A series of normal stages in the development of the chick embryo. J. Morphol. 88: Harvey, A. J., G. Speksnijder, L. R. Baugh, J. A. Morris, and R. Ivarie Expression of exogenous protein in the egg white of transgenic chickens. Nat. Biotechnol. 20: Jung, J. G., T. S. Park, J. N. Kim, B. K. Han, S. D. Lee, G. Song, and J. Y. Han Characterization and application of oviductal epithelial cells in vitro in Gallus domesticus. Biol. Reprod. 85: Kim, M. A., T. S. Park, J. N. Kim, H. J. Park, Y. M. Lee, T. Ono, J. M. Lim, and J. Y. Han Production of quail (Coturnix japonica) germline chimeras by transfer of gonadal primordial germ cells into recipient embryos. Theriogenology 63: Kwon, S. C., J. W. Choi, H. J. Jang, S. S. Shin, S. K. Lee, T. S. Park, I. Y. Choi, G. S. Lee, G. Song, and J. Y. Han Production of biofunctional recombinant human interleukin 1 receptor antagonist (rhil1rn) from transgenic quail egg white. Biol. Reprod. 82: Lillico, S. G., M. J. McGrew, A. Sherman, and H. M. Sang Transgenic chickens as bioreactors for protein-based drugs. Drug Discov. Today 10: Lillico, S. G., A. Sherman, M. J. McGrew, C. D. Robertson, J. Smith, C. Haslam, P. Barnard, P. A. Radcliffe,

7 191 K. A. Mitrophanous, E. A. Elliot, and H. M. Sang Oviduct-specific expression of two therapeutic proteins in transgenic hens. Proc. Natl. Acad. Sci. 104: Motono, M., Y. Yamada, Y. Hattori, R. Nakagawa, K. Nishijima, and S. Iijima Production of transgenic chickens from purified primordial germ cells infected with a lentiviral vector. J. Biosci. Bioeng. 109: Ono, T., R.Yokoi, and H. Aoyama Transfer of male or female primordial germ cells of quail into chick embryonic gonads. Exp. Anim. 45: Park, F Lentiviral vectors: are they the future of animal transgenesis? Physiol. Genomics 31: Park, T. S., H. G. Lee, J. K. Moon, H. J. Lee, J. W. Yoon, B. N. Yun, S. C. Kang, J. Kim, H. Kim, J. Y. Han, and B. K. Han Deposition of bioactive human epidermal growth factor in the egg white of transgenic hens using an oviduct-specific minisynthetic promoter. FASEB J. 29: Poynter, G., and R. Lansford Generating transgenic quail using lentiviruses. Methods Cell Biol. 87: Poynter, G., D. Huss, and R. Lansford Japanese quail: an efficient animal model for the production of transgenic avians. Cold Spring Harbor Protocols Poynter, G., D. Huss, and R. Lansford Injection of Lentivirus into Stage-X Blastoderm for the Production of Transgenic Quail. Cold Spring Harbor Protocols Scott, B. B., and C. Lois Generation of tissue-specific transgenic birds with lentiviral vectors. Proc. Natl. Acad. Sci. 102: Shin, S. S., T. M. Kim, S. Y. Kim, T. W. Kim, H. W. Seo, S. K. Lee, S. C. Kwon, G. S. Lee, H. Kim, J. M. Lim, and J. Y. Han Generation of transgenic quail through germ cell-mediated germline transmission. FASEB J. 22: Wyburn, G. M., H. S. Johnston, M. H. Draper, and M. F. Davidson The fine structure of the infundibulum and magnum of the oviduct of Gallus domesticus. Q. J. Exp. Physiol. Cogn. Med. Sci. 55:

8 192 J. Chin. Soc. Anim. Sci. 46(3): 185~192, 2017 Review Generation of transgenic poultry for producing functional proteins Ho-Ching Tsai (1) and Shih-Torng Ding (1)(2) ABSTRACT Transgenic animals have been used for producing functional proteins for decades. Compared with mammals, applications of transgenic poultry have attracted attentions because of short reproduction cycle and less space requirement. Eggs are produced by birds as a stable and reliable protein source. The simple composition and tissue-specific secretion of albumin makes it possible to accumulate target proteins only in egg white, and reduce the transgene affecting other tissues in the transgenic birds. The functional proteins produced by transgenic poultry are expected to be widely applied in different bio-medical fields. (Key Words: Functional proteins, Poultry, Transgenesis) (1) Department of Animal Science and Technology, National Taiwan University, No. 50, Lane 155, Sec. 3, Keelung Rd., Taipei 106, Taiwan. (2) Corresponding author: sding@ntu.edu.tw

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附件2：中国实验动物学会团体标准实验动物肿瘤坏死因子 α 转基因关节炎小鼠模型饲养繁育规范编制说明联系人 : 李舸联系电话 :15011983691 编制单位 : 广东省实验动物监测所时间 : 2016 年 10 月 25 日中国实验动物学会团体标准人肿瘤坏死因子 α 转基因关节炎小鼠模型饲养繁育规范编制说明 GB/T1.1 2009 1 2011A081401005 2013B040400007