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hEPO 유전자의 유선조직 특이적 발현에 대한 In Vitro 검정

In Vitro Assay of Mammary Gland Tissue Specific hEPO Gene Expression

  • 구본철 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 권모선 (대구가톨릭대학교 의과대학 생리학교실) ;
  • 김태완 (대구가톨릭대학교 의과대학 생리학교실)
  • Koo, Bon Chul (Department of Physiology, Catholic University of Daegu School of Medicine) ;
  • Kwon, Mo Sun (Department of Physiology, Catholic University of Daegu School of Medicine) ;
  • Kim, Teoan (Department of Physiology, Catholic University of Daegu School of Medicine)
  • 투고 : 2016.03.02
  • 심사 : 2016.03.07
  • 발행 : 2016.02.29

초록

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat ${\beta}$-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

키워드

참고문헌

  1. Ball RK, Friis RR, Schoenenberger CA, Doppler W, Groner B (1988): Prolactin regulation of beta-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line. EMBO J 7:2089-2095.
  2. Batista RI, Melo CH, Souza-Fabjan JM, Teixeira DI, Melo LM, Freitas VJ (2014): Phenotypic features of first-generation transgenic goats for human granulocyte-colony stimulation factor production in milk. Biotechnol Lett 36:2155-2162. https://doi.org/10.1007/s10529-014-1588-0
  3. Brophy B, Smolenski G, Wheeler T, Wells D, L' Huillier P, Laible G (2003): Cloned transgenic cattle produce milk with higher levels of beta-casein and kappa-casein. Nat Biotechnol 21:157-162. https://doi.org/10.1038/nbt783
  4. Burdon, TG, Maitland KA, Clark AJ, Wallace R, Watson CJ (1994): Regulation of the sheep beta-lactoglobulin gene by lactogenic hormones is mediated by a transcription factor that binds an interferongamma activation site-related element. Mol Endocrinol 8:1528-1536.
  5. Clark AJ (1998): The mammary gland as a bioreactor: expression, processing, and production of recombinant proteins. J Mammary Gland Biol Neoplasia 3:337-350. https://doi.org/10.1023/A:1018723712996
  6. Danielson KG, Oborn CJ, Durban EM, Butel JS, Medina D (1984): Epithelial mouse mammary cell line exhibiting normal morphogenesis in vivo and functional differentiation in vitro. Proc Natl Acad Sci USA 81:3756-3760. https://doi.org/10.1073/pnas.81.12.3756
  7. Doppler W, Groner B, Ball RK (1989): Prolactin and glucocorticoid hormones synergistically induce expression of transfected rat beta-casein gene promoter constructs in a mammary epithelial cell line. Proc Natl Acad Sci USA 86:104-108. https://doi.org/10.1073/pnas.86.1.104
  8. Doppler W, Villunger A, Jennewein P, Brduscha K, Groner B, Ball RK (1991): Lactogenic hormone ans cell-type specific control of the whey acidic protein gene promoter in transfected mouse cells. Mol Endocrinol 5:1624-1632. https://doi.org/10.1210/mend-5-11-1624
  9. Dyck MK, Lacroix D, Pothier F, Sirard MA (2003): Making recombinant proteins in animals - different systems, different applications. Trends Biotechnol 21: 394-399. https://doi.org/10.1016/S0167-7799(03)00190-2
  10. Fisher JW (2003): Erythropoietin: physiology and pharmacology update. Exp Biol Med 228:1-14. https://doi.org/10.1177/153537020322800101
  11. Fussenegger M, Bailey JE, Hauser H, Mueller PP (1999): Genetic optimization of recombinant glycoprotein production by mammalian cells. Trends Biotech 17:35-42. https://doi.org/10.1016/S0167-7799(98)01248-7
  12. Houdebine LM (2009): Production of pharmaceutical proteins by transgenic animals. Comp Immunol Microbiol Infect Dis 32:107-121. https://doi.org/10.1016/j.cimid.2007.11.005
  13. Jacobs C, Horl WH, Macdougallm IC (2000): European best practice guidelines 9-13: anaemia management. Nephrol Dial Transpl 15:33-42.
  14. Kim NY, Kim JH, Kim HJ (2005): Effect of low adapted temperature and medium composition on growth and erythropoietin (EPO) production by Chinese hamster ovary cells. Arch Pharm Res 28:220-226. https://doi.org/10.1007/BF02977719
  15. Ko JH, Lee CS, Kim KH, Pang MG, Koo JS, Fang N, Koo DB, Oh KB, Youn WS, Zheng GD, Park JS, Kim SJ, Han YM, Choi IY, Lim J, Shin ST, Jin SW, Lee KK, Yoo OJ (2000): Production of biologically active human granulocyte colony stimulating factor in the milk of transgenic goat. Transgenic Res 9: 215-222. https://doi.org/10.1023/A:1008972010351
  16. Krantz SB (1991): Erythropoietin. Blood 77:419-434.
  17. Levine JF, Stockdale FE (1985): Cell-cell interactions promote mammary epithelial cell differentiation. J Cell Biol 100:1415-1422. https://doi.org/10.1083/jcb.100.5.1415
  18. Lipinski D, Zeyland J, Szalata M, Plawski A, Jarmuz M, Jura J, Korcz A, Smorag Z, Pienkowski M, Slomski R (2012): Expression of human growth hormone in the milk of transgenic rabbits with transgene mapped to the telomere region of chromosome 7q. J Appl Genet 53:435-442. https://doi.org/10.1007/s13353-012-0110-4
  19. Maga EA, Shoemaker CF, Rowe JD, Bondurant RH, Anderson GB, Murray JD (2006): Production and processing of milk from transgenic goats expressing human lysozyme in the mammary gland. J Dairy Sci 89:518-524. https://doi.org/10.3168/jds.S0022-0302(06)72114-2
  20. Massoud M, Atta J, Thepot D, Pointu H, Stinnakre MG, Theron MC, Lopez C, Houdebine LM (1996): The deleterious effects of human erythropoietin gene driven by the rabbit whey acidic protein gene promoter in transgenic rabbits. Reprod Nutr Dev 36:555-563. https://doi.org/10.1051/rnd:19960511
  21. Maxwell AP (2002): Novel erythropoiesis stimulating protein in the management of the anemia of chronic renal failure. Kidney Int 62:720-729. https://doi.org/10.1046/j.1523-1755.2002.00474.x
  22. Meade HM, Echelard Y, Ziomek CA, Young MW, Harvey M, Cole ES, Groet S, Smith TE, Curling JM (1998): Expression of recombinant proteins in the milk of transgenic animals. In: Gene expression systems: using nature for the art of expression. Fernandez JM, Hoeffler JP (eds). Academic Press, San Diego, pp 399-427.
  23. McKnight RA, Shamay A, Sankaran L, Wall RJ, Hennighausen L (1992): Matrix attachment regions can impart position independent regulation of a tissue-specific gene in transgenic mice. Proc Natl Acad Sci USA 89:6943-6947. https://doi.org/10.1073/pnas.89.15.6943
  24. Mikus T, Maly P, Poplstein M, Landa V, Trefil P, Lidicky J (2001): Expression of human erythropoietin gene in the mammary gland of a transgenic mouse. Folia Biol (Praha) 47:187-195.
  25. Mikus T, Poplstein M, Sedlakova J, Landa V, Jenikova G, Trefil P, Lidicky J, Maly P (2004): Generation and phenotypic analysis of a transgenic line of rabbits secreting active recombinant human erythropoietin in the milk. Transgenic Res 13:487-498. https://doi.org/10.1007/s11248-004-9596-9
  26. Paleyanda RK1, Zhang DW, Hennighausen L, Mc-Knight RA, Lubon H (1994): Regulation of human protein C gene expression by the mouse WAP promoter. Transgenic Res 3:335-343. https://doi.org/10.1007/BF01976765
  27. Tong J, Wei H, Liu X, Hu W, Bi M, Wang Y, Li Q, Li N (2011): Production of recombinant human lysozyme in the milk of transgenic pigs. Transgenic Res 20:417-419. https://doi.org/10.1007/s11248-010-9409-2
  28. Topper YJ, Freeman CS (1980): Multiple hormone interactions in the developmental biology of the mammary gland. Physiol Rev 60:1049-1106. https://doi.org/10.1152/physrev.1980.60.4.1049
  29. van Berkel PH, Welling MM, Geerts M, van Veen HA, Ravensbergen B, Salaheddine M, Pauwels EK, Pieper F, Nuijens JH, Nibbering PH (2002): Large scale production of recombinant human lactoferrin in the milk of transgenic cows. Nat Biotechnol 20: 484-487. https://doi.org/10.1038/nbt0502-484
  30. Varki A, Cummings R, Esko J, Freeze H, Hart G, Marth J (1999): Essentials of Glycobiology. Cold Spring Harbor Laboratory Press, New York.
  31. Wu X, Lin Y, Xiong F, Zhou Y, Yu F, Deng J, Huang P, Chen H (2012): The extremely high level expression of human serum albumin in the milk of transgenic mice. Transgenic Res 21:1359-1366. https://doi.org/10.1007/s11248-012-9612-4
  32. Yang B, Wang J, Tang B, Liu Y, Guo C, Yang P, Yu T, Li R, Zhao J, Zhang L, Dai Y, Li N (2011): Characterization of bioactive recombinant human lysozyme expressed in milk of cloned transgenic cattle. PLoS One 16;6:e17593.
  33. Yekta AA, Dalman A, Eftekhari-Yazdi P, Sanati MH, Shahverdi AH, Fakheri R, Vazirinasab H, Daneshzadeh MT, Vojgani M, Zomorodipour A, Fatemi N, Vahabi Z, Mirshahvaladi S, Ataei F, Bahraminejad E, Masoudi N, Rezazadeh Valojerdi M, Gourabi H (2013): Production of transgenic goats expressing human coagulation factor IX in the mammary glands after nuclear transfer using transfected fetal fibroblast cells. Transgenic Res 22: 131-142. https://doi.org/10.1007/s11248-012-9634-y
  34. Yu H, Chen J, Liu S, Zhang A, Xu X, Wang X, Lu P, Cheng G (2013): Large-scale production of functional human lysozyme in transgenic cloned goats. J Biotechnol 168:676-83. https://doi.org/10.1016/j.jbiotec.2013.10.023