Expression Study of a Recombinant Plasmid containing Dipeptidyl Peptidase-4 Gene in E. coli: A Plausible Application for Celiac Disease Patients to Digest Gluten

  • Lee, Yeonjae (Hankuk Academy of Foreign Studies) ;
  • Kang, Ryan (Hankuk Academy of Foreign Studies) ;
  • Kwon, Jenna (Hankuk Academy of Foreign Studies) ;
  • Jo, Kyuhee (Hankuk Academy of Foreign Studies) ;
  • Im, Jungbin (Hankuk Academy of Foreign Studies) ;
  • Jung, Sangwook (Hankuk Academy of Foreign Studies) ;
  • Lee, DongHyun (Hankuk Academy of Foreign Studies) ;
  • Lee, Juhyeon (Hankuk Academy of Foreign Studies) ;
  • Lee, Jeong-Sang (Dep. of Biotechnology and Functional Foods, Jeonju University)
  • Received : 2018.05.15
  • Accepted : 2018.05.29
  • Published : 2018.06.30


Celiac disease (CD) is an immune-mediated enteropathy of small intestine diagnosed in both childhood and adulthood. CD is caused by gluten, which produces gliadorphin during its digestion. The enzyme dipeptidyl peptidase-4 (DPP4) breaks gliadorphin down nevertheless the last tripeptide remains and eventually inhibits DPP4, thus slowing down its process. Therefore, the idea is to produce an additional DPP4 enzyme which is crucial. Consequently, the functional DPP4 gene was cloned into pCDNA3 intermediate (FLAG+DPP4) vector and finally a recombinant plasmid pSB1C3 (Andersons promoters+FLAG+DPP4) was constructed using synthetic biology. Normally, a DPP4 inhibitor is used as a cure for diabetes. Another important concern was overexpression of DPP4, which might lead to diabetes, accordingly the work was also performed for the regulation of the DPP4 gene expression. In this regard, three types of Anderson promoters (strong, moderate and weak) were utilized to study the control overexpression. This is the first report of idealistic trial for control the exogenous DPP4 gene-expression by molecular biologic tools.


Celiac disease;Vectors;Anderson promoter;Infusion ligation;Synthetic biology


  1. J.M. Turner, "Diagnosis of Celiac Disease: Taking a Bite Out of the Controversy," Digestive Disease and Sciences (DDS), Vol. 63, No. 6, pp. 1384-1391 June, 2018. DOI: 10.1007/s10620-018-5050.
  2. J.T. Rosenbaum, "Celiac Disease and Autoimmunity-The Missing Ingredient," New England Journal of Medicine (NEJM), Vol. 377, No. 15, pp. 1489-1490, October, 2017. DOI: 10.1056/NEJMcibr1706917.
  3. J. West, K. M. Fleming, L. J. Tata, T. R. Card, and C. J. Crooks, "Incidence and Prevalence of Celiac Disease and Dermatitis Herpetiformis in the UK Over Two Decades: Population-Based Study," The American Journal Of Gastroenterology(AJG), Vol. 109, pp. 757-768, March, 2014. DOI: 10.1038/ajg.2014.55
  4. E. Almallouhi, K. S. King, B. Patel, C. Wi, Y. J. Juhn, J. A. Murray, and I. Absah, "Increasing Incidence and Altered Presentation in a Population-based Study of Pediatric Celiac Disease in North America," Journal of Pediatric Gastroenterology and Nutrition, Vol. 65, No. 4, pp. 432-437. October, 2017. DOI: 10.1097/MPG.0000000000001532.
  5. M.M. Leonard, A. Sapone, C. Catassi, and A. Fasano, "Celiac Disease and Nonceliac Gluten Sensitivity-a Review," JAMA, Vol. 318, No. 7, pp. 647-656, August, 2017. DOI:10.1001/jama.2017.9730.
  6. N.R. Lewis, and G.K.T. Holmes, "Risk of morbidity in contemporary celiac disease," Expert Review of Gastroenterology & Hepatology, Vol. 4, No. 6, pp. 767-780, December, 2010. DOI: 10.1586/egh.10.72.
  7. A. Sood, V. Midha, G. Makharia, B. K. Thelma, S. S. Halli, V. Mehta, R. Mahajan, V. Narang, K. Sood, and K. Kaur, "A simple phenotypic classification for celiac disease," Intestinal Research (IR), Vol. 16, No. 2, pp. 288-292, April, 2018. DOI: 10.5217/ir.2018.16.2.288.
  8. D.H. Dewar, and P.J. Ciclitira, "Clinical features and diagnosis of celiac disease," Gastroenterology (G), Vol. 128, No. 4, pp. 19-24, April, 2005.DOI:10.1053/j.gastro.2005.02.010.
  9. C. Klemann, L. Wagner, M. Stephan, and Horsten, "Cut to the chase: a review of CD26/dipeptidyl peptidase‐4's (DPP4) entanglement in the immune system. Clinical & Experimental Immunology (CEI), Vol. 185, No. 1, pp. 1-21, February, 2016. DOI: 10.1111/cei.12781.
  10. Y. Zhao, L. Yang, X. Wang, and Z. Zhou, "The New insights from DPP-4 inhibitors: their potential immune modulatory function in autoimmune diabetes, Diabetes/Metabolism Research and Reviews (DMRR), Vol. 30, No. 8, pp. 646-665, January, 2014. DOI: 10.1002/dmrr.2530
  11. September, 2015).
  12. J. Sambrook, and D.W. Russell, The Inoue Method for Preparation and Transformation of Competent E. Coli: "Ultra-Competent" Cells. Cold Spring Harbor Protocols, 2006. 2006(1): p. pdb.prot3944.
  13. J. Sambrook, E.F. Fritsch, and T. Maniatis, "Molecular cloning: a laboratory manual". 1989: Cold spring harbor laboratory press.
  14. T.P. Hopp, K.S. Prickett, V.L. Price, R.T. Libby, C.J. March, D.P. Cerretti, D.L. Urdal, and P.J. Conlon, "A short polypeptide marker sequence useful for recombinant protein identification and purification," Bio-Technology (BT), Vol. 6, No. 10, pp. 1204-1210, October, 1988. DOI: 10.1038/nbt1088-1204.
  15. J. F. Ludvigsson, T. Card, P. J. Ciclitira, G. L. Swift, I. Nasr, D. S Sanders and C. Ciacci, "Support for patients with celiac disease: A literature review," United European Gastroenterology Journal (UEG), Vol. 3, No. 2, pp. 146-159, April, 2015. DOI: 10.1177/2050640614562599.