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牛防御素BNBD4成熟肽的真核表达及纯化
Eukaryotic Expression and Purification of Mature Bovine Neutrophil β-Defensins 4

DOI: 10.12677/HJBM.2019.94025, PP. 169-176

Keywords: 防御素，BNBD4，成熟肽，真核表达，纯化
Defensins, BNBD4, Mature Peptide, Eukaryotic Expression, Purification

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Abstract:

牛β防御素4 (BNBD4)是一种天然存在于牛肺脏肺泡巨噬细胞内的抗菌肽，本研究拟通过克隆BNBD4成熟肽片段的编码区序列，连接至真核表达载体pPIC9K构建重组质粒mBNBD4-pPIC9K，转化至毕赤酵母菌株GS115诱导表达，镍柱纯化后，经Tricine-SDS-PAGE检测和Western Blot鉴定，成功获取重组蛋白mBNBD4，为进一步研究牛防御素BNBD4的作用机制提供试验材料。
Bovine neutrophil β-defensins 4 (BNBD4) is one of the antimicrobial peptides which naturally exists in bovine pulmonary alveolar macrophages. In this study we will clone the CDs sequence of mature BNBD4 gene, and connect it to the eukaryotic expression vector pPIC9K to construct recombinant plasmid mBNBD4-pPIC9K, and purify it by Ni-sepharose after inducing expression. Finally, we can successfully obtain the recombinant protein mBNBD4 by Tricine SDS-PAGE detection and Western Blot identify, therefore to provide the experimental materials for the further study of mature BNBD4.

References

[1]  Ganz, T. (2003) Defensins: Antimicrobial Peptides of Innate Immunity. Nature Reviews Immunology, 3, 710-720.
https://doi.org/10.1038/nri1180
[2]  Ganz, T., et al. (1985) Defensins. Natural Peptide Antibiotics of Human Neutrophils. Journal of Clinical Investigation, 76, 1427-1435.
https://doi.org/10.1172/JCI112120
[3]  Hi, Z. and Spitznagel, J.K. (1966) Cationic Proteins of Polymorphonuclear Leukocyte Lysosomes. I. Resolution of Antibacterial and Enzymatic Activities. Journal of Bacteriology, 91, 750-754.
[4]  Hi, Z. and Spitznagel, J.K. (1966) Cationic Proteins of Polymorphonuclear Leukocyte Lysosomes. II. Composition, Properties, and Mechanism of Antibacterial Action. Journal of Bacteriology, 91, 755-762.
[5]  Ri, L. and Ganz, T. (2002) Defensins of Vertebrate Animals. Current Opinion in Immunology, 14, 96-102.
https://doi.org/10.1016/S0952-7915(01)00303-X
[6]  Ganz, T. (2004) Defensins: Antimicrobial Peptides of Vertebrates. Comptes Rendus Biologies, 327, 539-549.
https://doi.org/10.1016/j.crvi.2003.12.007
[7]  Mendez-Samperio, P. (2008) Role of Antimicrobial Peptides in Host Defense against Mycobacterial Infections. Peptides, 29, 1836-1841.
https://doi.org/10.1016/j.peptides.2008.05.024
[8]  Shin, D.M. and Jo, E.K. (2011) Antimicrobial Peptides in Innate Immunity against Mycobacteria. Immune Network, 11, 245-252.
https://doi.org/10.4110/in.2011.11.5.245
[9]  Dong, H., Lv, Y., Zhao, D., Barrow, P. and Zhou, X.M. (2016) Defensins: The Case for Their Use against Mycobacterial Infections. Journal of Immunology Research, 2016, Article ID: 7515687.
https://doi.org/10.1155/2016/7515687
[10]  牛明福, 李翔, 邢广旭, 等. 6 × His-Tag在抗菌肽表达纯化中的应用及其对抗菌肽活性的影响[J]. 河南农业科学, 2008(12): 121-124.
[11]  Patoli, A.A. and Patoli, В.В. (2019) The N-Terminal 6 x His Tag on β-Clamp Processivity Factor Occludes Gly66 and Affects the Growth of Escherichia coli B834 (DE3) Cells. Molekuliarnaia Biologiia, 53, 32-37.
https://doi.org/10.1134/S0026893319010126
[12]  Schagger, H. (2006) Tricine-SDS-PAGE. Nature Protocols, 1, 16-22.
https://doi.org/10.1038/nprot.2006.4
[13]  Gasser, B., et al. (2013) Pichia pastoris: Protein Production Host and Model Organism for Biomedical Research. Future Microbiology, 8, 191-208.
https://doi.org/10.2217/fmb.12.133
[14]  Daly, R. and Hearn, M.T. (2005) Expression of Heterologous Proteins in Pichia pastoris: A Useful Experimental Tool in Protein Engineering and Production. Journal of Molecular Recognition, 18, 119-138.
https://doi.org/10.1002/jmr.687

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