Anti-Colorectal Cancer Effects of Probiotic-Derived p8 Protein
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains and Culture
2.2. Construction of Codon-Optimized His-Tagged r-p8 Protein, and Expression and Purification in E. coli
2.3. Expression of Codon-Optimized r-p8 Protein in DLD-1 Cells
2.4. Cell Culture
2.5. Cell Proliferation Assay
2.6. Wound Healing Assay
2.7. ELISA Analysis
2.8. Western Blot Analysis
2.9. Immunocytochemistry Using ImageXpress® Micro Confocal Microscopy
2.10. Flow Cytometry Analysis
3. Result
3.1. P8 Requires a Specific Delivery System
3.2. Endogenous p8 Expression Showed Markedly Enhanced Anti-Cancer Activity
3.3. Effects of r-p8 on Anti-Cancer Signaling Pathways in DLD-1 Cells
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Target Organism/Mapping | Codon-Optimized Sequences | Size (bp) |
---|---|---|
Original P8 | atggcaacagtagatcctgaaaagacattgtttctcgatgaaccaatgaacaaggtatttgactggagcaacagcgaagcacctgtacgtgatgcgctgtgggattattacatggaaaagaacagccgtgataccatcaagactgaagaagaaatgaaaccagtcctagacatgtccgacgatgaggtcaaagccctagcagaaaaggttctcaagaagtaa | 222 |
E. coli cells/6×His-TEV-P8 (NdeI/EcoRI) | catatgagaggatcgcatcaccatcaccatcac-attacgatatcccaacgaccgaaaacctgtattttcagggatcc-atggcaacagtagatcctgaaaagacattgtttctcgatgaaccaatgaacaaggtatttgactggagcaacagcgaagcacctgtccgtgatgcgctgtgggattattacatggaaaagaacagccgtgatactatcaagactgaagaagaaatgaaaccagtcctagacatgtccgacgacgaggtcaaagccctagcagaaaaggttctcaagaagtaggaattc | 305 |
DLD-1 cells/P8 (EcoRI/NotI) | gaattcatggctactgtcgacccagaaaaaaccctgttcttggacgaaccaatgaataaagtctttgattggtccaactctgaggccccggtacgggatgcgttgtgggattactacatggaaaaaaattccagggataccattaaaacagaagaagaaatgaagccagttctggacatgagtgacgacgaagtgaaagccctcgcggaaaaagttctcaagaaataggcggccgc | 236 |
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An, B.C.; Hong, S.; Park, H.J.; Kim, B.-K.; Ahn, J.Y.; Ryu, Y.; An, J.H.; Chung, M.J. Anti-Colorectal Cancer Effects of Probiotic-Derived p8 Protein. Genes 2019, 10, 624. https://doi.org/10.3390/genes10080624
An BC, Hong S, Park HJ, Kim B-K, Ahn JY, Ryu Y, An JH, Chung MJ. Anti-Colorectal Cancer Effects of Probiotic-Derived p8 Protein. Genes. 2019; 10(8):624. https://doi.org/10.3390/genes10080624
Chicago/Turabian StyleAn, Byung Chull, Sunwoong Hong, Ho Jin Park, Bong-Kyu Kim, Jun Young Ahn, Yongku Ryu, Jae Hyung An, and Myung Jun Chung. 2019. "Anti-Colorectal Cancer Effects of Probiotic-Derived p8 Protein" Genes 10, no. 8: 624. https://doi.org/10.3390/genes10080624
APA StyleAn, B. C., Hong, S., Park, H. J., Kim, B.-K., Ahn, J. Y., Ryu, Y., An, J. H., & Chung, M. J. (2019). Anti-Colorectal Cancer Effects of Probiotic-Derived p8 Protein. Genes, 10(8), 624. https://doi.org/10.3390/genes10080624