Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (13): 2737-2742.doi: 10.3864/j.issn.0578-1752.2012.13.018

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Breeding and Expression Stability of Human Lactoferrin Transgenic Goat

 QIAO  Na, XU  Xu-Jun, LIU  Si-Guo, WANG  Xue-Bin, YU  Hui-Qing, TONG  Yong-Xing, CHEN  Jian-Quan, CHENG  Guo-Xiang   

  1. 1.上海转基因研究中心，上海 201210
    2.上海杰隆生物工程股份有限公司，上海 201210
  • Received:2011-10-31 Online:2012-07-01 Published:2012-04-10

PDF

809

Cited

2

Abstract: 【Objective】 This study was conducted to analyze the effects of transgenosis on the reproduction of human lactoferrin (hLF) transgenic goat, and the genetic and expression stability of exogenous gene in each generation of transgenic animals.【Method】Family line was established with self-developed hLF transgenic goats (G0♂), F1 generation was composed by integrated offspring individuals of G0♂ mated with ordinary dairy goat , F2 was generated by F1 males mated with F1, F2 or ordinary females, and F3 was generated by F2 males mated with F2 females. PCR and ELISA were implied to identify the integration and expression of exogenous gene in offsprings. Reproductive data such as quantity of offspring with different mating patterns, rate of transgenic integrated individuals, conception rates, abnormal rates of lamb in each generation were counted statistically. 【Result】 After several reproductive cycles, no abnormalities of conception rates and lambing rates were observed. The results showed that, there were 19 hLF transgenic individuals in F1 generation, with integration rate of 24.7% (19/77), 86 individuals in F2 generation, in which 62 were generated by F1♂×N♀, 16 were obtained by F1♂×F1♀, and 8 were obtained by F1♂×F2♀, with the rate of 33.2% (62/187), 76.2% (16/21) and 34.8% (8/23), respectively, 9 hLF transgenic individuals in F3 generation and the integration rate of F3 was 52.9% (9/17). ELISA was conducted to measure the expression of hLF in F1, F2 and F3 in 20 weeks, and the results showed that hLF expressed stably in each transgenic generation and the average expression level in milk were 351.75, 340.00 and 326.25 μg•mL-1, respectively.【Conclusion】The results suggested that, there is no significant effect of transgenosis on reproductive performance of hLF transgenic goat, and exogenous hLF gene could inherit and stably express in each generation of goats.

Key words: human lactoferrin, transgenic goat, breeding, hereditary, expression stability

[1]陈莎莎, 丁月云, 金  融, 王  恬. 乳铁蛋白的抑菌作用及其机制. 中国乳品工业, 2007, 35(5): 53-58.

Chen S S, Ding Y Y, Jin R, Wang T. Antibacterial function of lactoferrin (LF) and its mechanism. China Dairy Industry, 2007, 35(5): 53-58. (in Chinese)

[2]Gordon K, Lee E, Vitale J A, Smith A E, Westphal H, Hennighausen L. Production of human tissue plasminogen activator in transgenic mouse milk. Nature Biotechnology, 1987, 5: 1183-1187.

[3]Patrick H C, van B, Mick M W, Marlieke G, Harry A V, Bep R, Mourad S, Ernest K J, Frank P, Jan H N, Peter H, Nibbering. Large scale production of recombinant human lactoferrin in the milk of transgenic cows. Nature Biotechnology, 2002, 20: 484-487.

[4]Weinberg E D. Human lactoferrin: a novel therapeutic with broad spectrum potential. Journal of Pharmacy and Pharmacology, 2001, 53(10): 1303-1310.

[5]Brundige D R, Maga E A, Klasing K C, Murray J D. Lysozyme transgenic goats’ milk influences gastrointestinal morphology in young pigs. Nutrition, 2008, 138(5): 921-926.

[6]Scharfen E C, Mills D A, Maga E A. Use of human lysozyme transgenic goat milk in cheese making: effects on lactic acid bacteria performance. American Dairy Science Association, 2007, 90(9): 4084-4091.

[7]Maga E A, Cullor J S, Smith W, Anderson G B, Murray J D. Human lysozyme expressed in the mammary gland of transgenic dairy goats can inhibit the growth of bacteria that cause mastitis and the cold-spoilage of milk. Foodborne Pathogens and Disease, 2006, 3(4): 384-392.

[8]支旭勃, 李青旺, 冯  波, 肖  波, 韩增胜, 范玉生, 张  婷. 转基因羊乳中重组人乳铁蛋白的分离工艺研究. 黑龙江畜牧兽医, 2008(5): 83-84.

Zhi X B, Li Q W, Feng B, Xiao B, Han Z S, Fan Y S, Zhang T. Study on the separation process for recombinant human lactoferrin transgenic goat milk. Heilongjiang Animal Science and Veterinary Medicine, 2008(5): 83-84. (in Chinese)

[9]Maga E A, Walker R L, Anderson G B, Murray J D. Consumption of milk from transgenic goats expressing human lysozyme in the mammary gland results in the modulation of intestinal microflora. Trasgenic Research, 2006, 15(4): 515-519.

[10]Maga E A, Shoemaker C F, Rowe J D, Bondurant R H, Anderson G B, Murry J D. Production and processing of milk from transgenic goats expressing human lysozyme in the mammary gland. American Dairy Science Association, 2006, 89(2): 518-524.

[11]Krimpenfort P. The production of human lactoferrin in the milk of transgenic animals. Cancer Detection and Prevention, 1993, 17(2): 301-305.

[12]James W L, David W T. Producing proteins in transgenic plants and animals. Current Opinion in Biotechnology, 2001, 12(4): 411-418.

[13]Yang P H, Wang J W, Gong G C, Sun X Z, Zhang R, Du Z, Liu Y, Li R, Ding F R, Tang B, Dai Y P, Li N. Cattle mammary bioreactor generated by a novel procedure of transgenic cloning for large-scale production of functional human lactoferrin. PLoS One, 2008, 3(10): e3453.

[14]李佐刚, 汤  瑶, 孙  旭, 闻  镍, 于  敏, 史兴昌, 李  响, 王秀文, 王军志, 李  波. 转基因山羊羊乳中重组人乳铁蛋白性质的检定与研究. 药物分析杂志, 2008, 28(2): 223-226.

Li Z G, Tang Y, Sun X, Wen N, Yu M, Shi X C, Li X, Wang X W, Wang J Z, Li B. Identification and characterization of recombinant human lactoferrin in the milk of transgenic goats. Chinese Journal of Pharmaceutical Analysis, 2008, 28(2): 223-226. (in Chinese)

[15]张  然, 王媛媛, 鲍永华, 赵　杰,　李  宁. 转基因动物应用的研究现状与生物安全评价. 生物产业技术, 2010, 3: 48-59.

Zhang R, Wang Y Y, Bao Y H, Zhao J, Li N. Applications of transgenic animal research status and evaluation of biosafety. Biotechnology and Business, 2010, 3: 48-59. (in Chinese)

[16]Kathryn A J, Jolene M B, James D M, Elizabeth A M. Evaluating the fitness of human lysozyme transgenic dairy goats: growth and reproductive traits. Trasgenic Research, 2010, 19: 977-986.

[17]王二先, 徐旭俊, 苗晋锋, 陈建泉, 成国祥, 杨  倩. 转人乳铁蛋白基因山羊乳对大鼠生长及代谢机能的影响. 南京农业大学学报, 2011, 34(2): 119-123.

Wang E X, Xu X J, Miao J F, Chen J Q, Cheng G X, Yang Q. Effect of transgenic human lactoferrin goats’ milk on growth and metabolic performance in rats. Journal of Nanjing Agricultural University, 2011, 34(2): 119-123. (in Chinese)

[18]Muir W M, Howard R D. Assessment of possible ecological risks and hazards of transgenic fish with implications for other sexually reproducing organisms. Trasgenic Research, 2002, 11: 101-114.

[19]赵有璋. 羊生产学. 北京：中国农业出版社, 2002.

Zhao Y Z. Sheep and Goat Production. Beijing: China Agricultural Press, 2002. (in Chinese)

[20]Kim S J, Yu D Y, Pak K W, Jeong S, Kim S W, Lee K K. Structure of the human lactoferrin gene and its chromosomal localization. Molecular and Cellular Proteomics, 1998, 8: 663-668

[21]Robert M B, Theresa K. Lactoferrin content of peripheral blood cells. British Journal of Haematology, 1978, 39(4): 509-521. 

[22]柏亚军. 人乳铁蛋白转基因小鼠遗传特性及rhLF抑菌活性分析[D]. 扬州: 扬州大学, 2008.

Bai Y J. Study on genetic characterization of transgenic mice and antibacterial activity of rhLF in the milk[D]. Yangzhou: Yangzhou University, 2008. (in Chinese)

[23]肖艳萍, 奚  鹰, 黄文英, 黄  英. 应用荧光原位杂交检测人凝血因子Ⅸ在转基因小鼠染色体上的整合. 遗传, 2002, 24(3): 232-236.

Xiao Y P, Xi Y, Huang W Y, Huang Y. Detection of the integration of human FⅨ on chromosomes of transgenic mice by fluorescence in situ hybridization. Hereditas, 2002, 24(3): 232-236. (in Chinese)

[24]刘祖洞. 遗传学. 北京: 高等教育出版社, 2006.

Liu Z D. Genetics. Beijing: Higher Education Press, 2006. (in Chinese)

[25]Falconer D S, Mackay T F. 储明星译. 数量遗传学导论. 第四版. 北京: 中国农业科技出版社, 2000.

Falconer D S, Mackay T F. Translated by Chu M X. Introduction to Quantitative Genetics. 4 ed. Beijing: China Agricultural Science and Technology Press, 2000. (in Chinese)

[26]华志华, 黄大年. 转基因植物中外源基因的遗传学行为. 植物学 报, 1999, 41(1): 1-5.

Hua Z H, Huang D N. Genetic mode of exogenes in transgenic plants. Acta Botanica Sinica, 1999, 41(1): 1-5. (in Chinese)

[27]Houdebine L M. The methods to generate transgenic animals and to control transgene expression. Biotechnology, 2002, 98: 145-160.

[28]Yamauchi Y, Doe B, Ajduk A, Ward M A. Genomic DNA damage in mouse transgenesis. Biology of Reproduction, 2007, 77: 803-812.

[29]朱士恩. 21世纪哺乳动物胚胎生物工程发展趋向. 中国农业科学, 2008, 41(8): 2425-2430.

Zhu S E. Trends in mammalian embryo bioengineering in the 21st century. Scientia Agricultura Sinica, 2008, 41(8): 2425-2430. (in Chinese)

[30]王  令, 张存芳, 张智英. 锌指核酸酶在基因组靶向修饰中的应 用. 中国生物化学与分子生物学报, 2009, 25(7): 585-589.

Wang L, Zhang C F, Zhang Z Y. Application of zinc finger nucleases in genome targeting modification. Chinese Journal of Biochemistry and Molecular Biology, 2009, 25(7): 585-589. (in Chinese)
[1] WANG CaiXiang,YUAN WenMin,LIU JuanJuan,XIE XiaoYu,MA Qi,JU JiSheng,CHEN Da,WANG Ning,FENG KeYun,SU JunJi. Comprehensive Evaluation and Breeding Evolution of Early Maturing Upland Cotton Varieties in the Northwest Inland of China [J]. Scientia Agricultura Sinica, 2023, 56(1): 1-16.
[2] LIU Shuo,ZHANG Hui,GAO ZhiYuan,XU JiLi,TIAN Hui. Genetic Variations of Potassium Harvest Index in 437 Wheat Varieties [J]. Scientia Agricultura Sinica, 2022, 55(7): 1284-1300.
[3] WANG Kai,ZHANG HaiLiang,DONG YiXin,CHEN ShaoKan,GUO Gang,LIU Lin,WANG YaChun. Definition and Genetic Parameters Estimation for Health Traits by Using on-Farm Management Data in Dairy Cattle [J]. Scientia Agricultura Sinica, 2022, 55(6): 1227-1240.
[4] MA HongXiang, WANG YongGang, GAO YuJiao, HE Yi, JIANG Peng, WU Lei, ZHANG Xu. Review and Prospect on the Breeding for the Resistance to Fusarium Head Blight in Wheat [J]. Scientia Agricultura Sinica, 2022, 55(5): 837-855.
[5] FENG XuanJun, PAN LiTeng, XIONG Hao, WANG QingJun, LI JingWei, ZHANG XueMei, HU ErLiang, LIN HaiJian, ZHENG HongJian, LU YanLi. Investigation on Important Target Traits and Breeding Potential of 120 Sweet and Waxy Maize Inbred Lines in the South of China [J]. Scientia Agricultura Sinica, 2022, 55(5): 856-873.
[6] LU Xiang, GAO Yuan, WANG Kun, SUN SiMiao, LI LianWen, LI HaiFei, LI QingShan, FENG JianRong, WANG DaJiang. Analysis of Aroma Characteristics in Different Cultivated Apple Strains [J]. Scientia Agricultura Sinica, 2022, 55(3): 543-557.
[7] JIANG Peng, ZHANG Peng, YAO JinBao, WU Lei, HE Yi, LI Chang, MA HongXiang, ZHANG Xu. Phenotypic Characteristics and Related Gene Analysis of Ningmai Series Wheat Varieties [J]. Scientia Agricultura Sinica, 2022, 55(2): 233-247.
[8] CHEN XueSen,WANG Nan,ZHANG ZongYing,MAO ZhiQuan,YIN ChengMiao. Understanding and Thinking About Some Problems of Fruit Tree Germplasm Resources and Genetic Breeding [J]. Scientia Agricultura Sinica, 2022, 55(17): 3395-3410.
[9] FANG TaoHong,ZHANG Min,MA ChunHua,ZHENG XiaoChen,TAN WenJing,TIAN Ran,YAN Qiong,ZHOU XinLi,LI Xin,YANG SuiZhuang,HUANG KeBing,WANG JianFeng,HAN DeJun,WANG XiaoJie,KANG ZhenSheng. Application of Yr52 Gene in Wheat Improvement for Stripe Rust Resistance [J]. Scientia Agricultura Sinica, 2022, 55(11): 2077-2091.
[10] JiaJia LI,HuiLong HONG,MingYue WAN,Li CHU,JingHui ZHAO,MingHua WANG,ZhiPeng XU,Yin ZHANG,ZhiPing HUANG,WenMing ZHANG,XiaoBo WANG,LiJuan QIU. Construction and Application of Detection Model for the Chemical Composition Content of Soybean Stem Based on Near Infrared Spectroscopy [J]. Scientia Agricultura Sinica, 2021, 54(5): 887-900.
[11] GAO ZhiYuan,XU JiLi,LIU Shuo,TIAN Hui,WANG ZhaoHui. Variations of Winter Wheat Nitrogen Harvest Index in Field Wheat Population [J]. Scientia Agricultura Sinica, 2021, 54(3): 583-595.
[12] ZHANG PengFei,SHI LiangYu,LIU JiaXin,LI Yang,WU ChengBin,WANG LiXian,ZHAO FuPing. Advance in Genome-Wide Scan of Runs of Homozygosity in Domestic Animals [J]. Scientia Agricultura Sinica, 2021, 54(24): 5316-5326.
[13] ZHANG Xingping,QIAN Qian,ZHANG JiaNan,DENG XingWang,WAN JianMin,XU Yunbi. Transforming and Upgrading Off-Season Breeding in Hainan Through Molecular Plant Breeding [J]. Scientia Agricultura Sinica, 2021, 54(18): 3789-3804.
[14] MENG JunRen,ZENG WenFang,DENG Li,PAN Lei,LU ZhenHua,CUI GuoChao,WANG ZhiQiang,NIU Liang. Development and Application of KASP Molecular Markers of Some Important Traits for Peach [J]. Scientia Agricultura Sinica, 2021, 54(15): 3295-3307.
[15] Xiao ZHANG,Man LI,DaTong LIU,Wei JIANG,Yong ZHANG,DeRong GAO. Analysis of Quality Traits and Breeding Inspiration in Yangmai Series Wheat Varieties [J]. Scientia Agricultura Sinica, 2020, 53(7): 1309-1321.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd