Expression of goat RNA binding protein (NANOS2), development and evaluation of anti-NANOS2 IgY antibodies X., Ren H., Yang W., Diraviyam T., Tang F., Yu M., Hua J., Zhang X., Peng S. (2015): Expression of goat RNA binding protein (NANOS2), development and evaluation of anti-NANOS2 IgY antibodies. Czech J. Anim. Sci., 60: 152-163.
download PDF

Available rabbit anti-NANOS2 antibodies failed to recognize the NANOS2 protein of goat origin though they showed better reactivity towards the same protein of human and mice origin. A specific antibody against goat NANOS2 is necessary to explore the biological functions of dairy goat NANOS2 protein. Recombinant NANOS2 protein (dairy goat origin) was expressed in E. coli (BL21). Anti-NANOS2 immunoglobulin Y (IgY; in chicken) was developed and evaluated. NANOS2 protein sequences between mice and dairy goat were compared using bioinformatics tools. The specific IgY titer attained the peak (≥ 1 : 128 000) after the 3rd booster injection, then it gradually decreased to 1 : 16 000 and remained stable. The specificity and sensitivity of anti-NANOS2 IgY was superior to rabbit anti-NANOS2 IgG. Though, the functional domain 63–116 or 62–115 of NANOS2 protein is highly conserved with 100% identity in dairy goat and mice, the considerable difference in sequence at region 18–61 was identified from the predicted epitopes. According to our survey, this is the first report on preparation of chicken IgY against NANOS2 protein and indicating the prerequisite of species specific NANOS2 antibodies due to epitopes difference in functionally conserved NANOS2 protein. Anti-NANOS2 IgY could be applied as a novel research tool in developmental biology.

An X.P., Hou J.X., Wang L.X., Li G., Wang J.G., Song Y.X., Zhou G.Q., Han D., Ling L., Cao B.Y. (2010): Novel polymorphisms of the growth hormone gene and their effect on growth traits in Chinese goats. Meat Science, 86, 758-763
Cao H., Chu Y., Zhu H., Sun J., Pu Y., Gao Z., Yang C., Peng S., Dou Z., Hua J. (2011): Characterization of immortalized mesenchymal stem cells derived from foetal porcine pancreas. Cell Proliferation, 44, 19-32
Dias da Silva Wilmar, Tambourgi Denise V. (2010): IgY: A promising antibody for use in immunodiagnostic and in immunotherapy. Veterinary Immunology and Immunopathology, 135, 173-180
Ding W., Kou L., Cao B., Wei Y. (2010): Meat quality parameters of descendants by grading hybridization of Boer goat and Guanzhong Dairy goat. Meat Science, 84, 323-328
Draper Bruce W., McCallum Claire M., Moens Cecilia B. (2007): nanos1 is required to maintain oocyte production in adult zebrafish. Developmental Biology, 305, 589-598
Hädge D., Ambrosius H. (1984): Evolution of low molecular weight immunoglobulins—IV. IgY-like immunoglobulins of birds, reptiles and amphibians, precursors of mammalian IgA. Molecular Immunology, 21, 699-707
Hardin C.C., Pinczes J., Riell A., Presutti D., Miller W., Robertson D. (eds) (2001): Cloning, Gene Expression, and Protein Purification. Oxford University Press, New York, USA.
Hu P. F., Guan W. J., Li X. C., Ma Y. H. (2012): Construction of recombinant proteins for reprogramming of endangered Luxi cattle fibroblast cells. Molecular Biology Reports, 39, 7175-7182
Hua Jinlian, Zhu Haijing, Pan Shaohui, Liu Chao, Sun Junwei, Ma Xiaoling, Dong Wuzi, Liu Weishuai, Li Wei (2011): Pluripotent Male Germline Stem Cells from Goat Fetal Testis and Their Survival in Mouse Testis. Cellular Reprogramming (Formerly "Cloning and Stem Cells"), 13, 133-144
KONNO A, MARICATO J, KONNO F, MARIANO M, LOPES J (2009): Peptides from Paracoccidioides brasiliensis GP43 inhibit macrophage functions and inflammatory response. Microbes and Infection, 11, 92-99
Kovacs-Nolan Jennifer, Mine Yoshinori (2004): Avian egg antibodies: basic and potential applications. Avian and Poultry Biology Reviews, 15, 25-46
LARSSON A., BALOW R.-M., LINDAHL T. L., FORSBERG P.-O. (1993): Chicken Antibodies: Taking Advantage of Evolution--A Review. Poultry Science, 72, 1807-1812
Li M., Liu C., Zhu H., Sun J., Yu M., Niu Z., Liu W., Peng S., Hua J. (2013a): Expression pattern of Boule in dairy goat testis and its function in promoting the meiosis in male germline stem cells (mGSCs). Journal of Cellular Biochemistry, 114, 294–302.
Li M., Yu M., Liu C., Zhu H., He X., Peng S., Hua J. (2013b): miR-34c works downstream of p53 leading to dairy goat male germline stem-cell (mGSCs) apoptosis. Cell Proliferation, 46, 223–231.
Li M., Yu M., Liu C., Zhu H., Hua J. (2013c): Expression of miR-34c in response to overexpression of Boule and Stra8 in dairy goat male germ line stem cells (mGSCs). Cell Biochemistry and Function, 31, 281–288.
Li M., Yu M., Zhu H., Song W., Hua J. (2013d): The effects of Nanos2 on Boule and Stra8 in male germline stem cells (mGSCs). Molecular Biology Reports, 40, 4383–4389.
Liu C., Zhu H., Liu W., Zheng W., Wang J., Yang C., Hua J. (2011): Male germ cells specification of embryonic gonad from Guanzhong dairy goat. Chinese Journal of Animal and Veterinary Sciences, 42, 1328–1336.
Mu Hailong, Wu Jiang, Zhu Haijing, Li Na, Tang Furong, Yao Xi, Yang Churong, Peng Sha, Li Guangpeng, Hua Jinlian (2013): The function of Msx1 gene in promoting meiosis of dairy goat male germline stem cells (mGSCs). Cell Biochemistry and Function, 31, 629-635
Olivier J.J., Cloete S.W.P., Schoeman S.J., Muller C.J.C. (2005): Performance testing and recording in meat and dairy goats. Small Ruminant Research, 60, 83-93
Orwig K.E., Hermann B.P. (eds) (2011): Male Germline Stem Cells: Development and Regenerative Potential. Humana Press, London, UK.
Polson A., von Wechmar M. Barbara, van Regenmortel M. H.V. (1980): Isolation of Viral IgY Antibodies from Yolks of Immunized Hens. Immunological Investigations, 9, 475-493
Qi Yu, Luo Jun, Han XueFeng, Zhu YanZhi, Chen Chao, Liu JunXia, Sheng HeJun (2009): Genetic diversity and relationships of 10 Chinese goat breeds in the Middle and Western China. Small Ruminant Research, 82, 88-93
Sambrook J., Russell D.W. (2001): Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, New York, USA.
Schade R., Hlinak A. (1995): Egg yolk antibodies, state of the art and future prospects. Altex, 13, 5–9.
Song W., Zhu H., Li M., Li N., Wu J., Mu H., Yao X., Han W., Liu W., Hua J. (2013): Promyelocytic leukaemia zinc finger maintains self-renewal of male germline stem cells (mGSCs) and its expression pattern in dairy goat testis. Cell Proliferation, 46, 457-468
Suzuki A., Saga Y. (2008): Nanos2 suppresses meiosis and promotes male germ cell differentiation. Genes & Development, 22, 430-435
Suzuki Hitomi, Sada Aiko, Yoshida Shosei, Saga Yumiko (2009): The heterogeneity of spermatogonia is revealed by their topology and expression of marker proteins including the germ cell-specific proteins Nanos2 and Nanos3. Developmental Biology, 336, 222-231
Tang F., Yao X., Zhu H., Mu H., Niu Z., Yu M., Yang C., Peng S., Li G., Hua J. (2014): Expression pattern of Ngn3 in dairy goat testis and its function in promoting meiosis by upregulating Stra8. Cell Proliferation, 47, 38-47
Wang Z. (2004): Nanos Maintains Germline Stem Cell Self-Renewal by Preventing Differentiation. Science, 303, 2016-2019
Xu Yongping, Li Xiaoyu, Jin Liji, Zhen Yuhong, Lu Yanan, Li Shuying, You Jiansong, Wang Linhui (2011): Application of chicken egg yolk immunoglobulins in the control of terrestrial and aquatic animal diseases: A review. Biotechnology Advances, 29, 860-868
Yao X., Tang F., Yu M., Zhu H., Chu Z., Li M., Liu W., Hua J., Peng S. (2014): Expression profile of Nanos2 gene in dairy goat and its inhibitory effect on Stra8 during meiosis. Cell Proliferation, 47, 396-405
Zhu Haijing, Liu Chao, Li Mingzhao, Sun Junwei, Song Wencong, Hua Jinlian (2013): Optimization of the conditions of isolation and culture of dairy goat male germline stem cells (mGSC). Animal Reproduction Science, 137, 45-52
Zhu Haijing, Ma Jing, Du Rui, Zheng Liming, Wu Jiang, Song Wencong, Niu Zhiwei, He Xin, Du Enqi, Zhao Shanting, Hua Jinlian (2014): Characterization of Immortalized Dairy Goat Male Germline Stem Cells (mGSCs). Journal of Cellular Biochemistry, 115, 1549-1560
Zhu Qinghe, Guo Donghua, Feng Li, Sun Dongbo (2013): Expression and Purification of the scFv from Hybridoma Cells Secreting a Monoclonal Antibody Against S Protein of PEDV. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, 32, 41-46
download PDF

© 2019 Czech Academy of Agricultural Sciences