Difference between revisions of "Part:BBa K4004002"

Revision as of 09:40, 20 October 2021

pMD19-T vp1

promoter

Profile

Name: pMD19-T vp1

Base Pairs: 3583bp

Origin: synthetic

Properties: Hand-foot-mouth disease Drinkable EV71 Vaccine

Usage and Biology

Hand-foot-mouth disease (HFMD) is an infectious disease caused by enterovirus 71 (EV71). The virus is an important pathogenic factor of hand, foot and mouth disease. Vp1 protein is the viral capsid protein and promotes the infection of host cells by virus particles. Vp1 is also the main antigen gene of the EV71 virus. Generally, the vaccinated population, especially infants and young children, are more compliant with oral vaccines, so we are trying to develop oral HFMD vaccines. Probiotics bacteria Bifidobacteria, as the natural host of the intestinal tract, can adhere to intestinal epithelial cells and are ideal oral live vaccine expression vectors, and related studies have found that their preventive effects on gastrointestinal pathogens are more significant. Therefore, we can use the bifidobacterium in lactic acid bacteria as an expression system to express EV71 vp1.

Construct design

This part is a recombinant plasmid with vp1 inserted. The design and schematic map are shown in Figure 1 and 2.

Figure 1 pMD19-T-vp1 box...

Figure 2. Schematic map of VP1 plasmid...

The profiles of every basic part are as follows:

BBa_K4004001

Name: vp1

Base Pairs: 891bp

Origin: E. coli

Properties: Vp1 is also the main antigen gene of the EV71 virus

Usage and Biology

BBa_K4004001 is a coding sequence of from E. coli . Vp1 protein is the viral capsid protein and promotes the infection of host cells by virus particles. Vp1 is also the main antigen gene of the EV71 virus.

BBa_K4004000

Name: pMD 19-T vector

Base Pairs: 2692bp

Origin: Synthetic

Properties: A plasmid dedicated for highly efficient cloning of PCR products.

Usage and Biology

BBa_K4004000 is a plasmid dedicated for highly efficient cloning of PCR products. This plasmid is reconstructed from pUC57 plasmid.

Experimental approach

Firstly, to amplify VP1 fragments from pUC57-VP1 and LTB fragments from pUC57-LTB, we added VP1-FP and VP1-RP into two tubes to amplify VP1 fragments.

Figure 3 Gel electrophoresis of VP1...

Conclusion: Theoretically, VP1 fragment is 891bp in length,we obtain it.

Then we performed enzyme digestion on vp1 and plasmid pMD 19-T. We could get pMD19-T backbone and vp1 fragment. We connected them with T4 ligase and finally we got recombinant plasmid pMD19-T-vp1.

References

1. Buch MH, Liaci AM, O'Hara SD, Garcea RL, Neu U, Stehle T (October 2015). "Structural and Functional Analysis of Murine Polyomavirus Capsid Proteins Establish the Determinants of Ligand Recognition and Pathogenicity". PLoS Pathogens. 11 (10): e1005104. doi:10.1371/journal.ppat.1005104

2. Ramqvist T, Dalianis T (August 2009). "Murine polyomavirus tumour specific transplantation antigens and viral persistence in relation to the immune response, and tumour development". Seminars in Cancer Biology. 19 (4): 236–43. doi:10.1016/j.semcancer.2009.02.001

3. Nassef, C., Ziemer, C., & Morrell, D. S. (2015). Hand-foot-and-mouth disease: a new look at a classic viral rash. Current opinion in pediatrics, 27(4), 486–491. https://doi.org/10.1097/MOP.0000000000000246

4. Who.int. 2021. How do vaccines work?. [online] Available at: <https://www.who.int/news-room/feature-stories/detail/how-do-vaccines-work?gclid=EAIaIQobChMIn4OC7YOh8gIVsG1vBB0wYgcmEAAYAyAAEgIBFvD_BwE> [Accessed 8 August 2021].

5. Yee, Pinn & Poh, Chit. (2015). Development of Novel Vaccines against Enterovirus-71. Viruses. 8. 1. 10.3390/v8010001.

6. Orlando, A.; Refolo, M. G.; Messa, C.; Amati, L.; Lavermicocca, P.; Guerra, V.; Russo, F. (October 2012). "Antiproliferative and Proapoptotic Effects of Viable or Heat-Killed IMPC2.1 and GG in HGC-27 Gastric and DLD-1 Colon Cell Lines". Nutrition and Cancer. 64 (7): 1103–1111. doi:10.1080/01635581.2012.717676

Sequence and Features