원문정보
초록
영어
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.
목차
1. Introduction
2. Theory
3. Experiments
3.1 PCR amplification of DPP4 gene from SPORT6 and purification
3.2 Preparation of the intermediate vector (pCDNA3) and Infusion ligation
3.3 Transformation and Plasmid isolation
3.4 Construction of backbone plasmid (pCDNA+FLAG+DPP4) and amplification
3.5 Transformation using pSB1C3
3.6 Infusion Ligation
3.7 Transfection
3.8 Western Blot analysis
4. Results and Discussion
4.1 Transformation of recombinant plasmid and infusion cloning
4.2 Expression of DPP4 protein
5. Conclusion
Acknowledgement
References