Analysis of genetic polymorphism in genes encoding cathelicidins from Vietnam indigenous yellow cattle breeds
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https://doi.org/10.15625/2615-9023/18295Keywords:
CATHL2, CATHL4, cathelicidins, genetic polymorphism, Vietnam indigenous cattle.Abstract
Cathelicidins include antimicrobial peptides (AMPs) and are involved in the innate immune defense against infections in mammals. Polymorphisms in the DNA sequence of cathelicidin genes could be relevant to inherited variations of host innate immunity, adaptation, and pathogen resistance. This study aims to investigate the sequence polymorphism of cathelicidin genes including CATHL2 and CATHL4 from local indigenous yellow cattle breeds of Vietnam. Genomic DNA samples were extracted from 52 individuals collected from different cattle populations in Vietnam including Ha Giang, Thanh Hoa, Nghe An and Phu Yen. The CATHL2 and CATHL4 genes were amplified by PCR, following sequencing for identification of the single nucleotide polymorphisms (SNPs) and/or insertion-deletion (indel) DNA sequence variations. The copy number variations (CNVs) of the CATHL4 gene were determined by cloning, single strand conformation polymorphism (SSCP) and real-time PCR. The comparative analysis results showed that there are 13 SNPs detected in all sequences of CATHL2 gene including 9 SNPs in intron 2 and intron 3, and 4 missense substitutions in exon 1, 2, and 3. These SNPs do not affect the predicted tertiary structure of Bac5 encoded by this gene. For CATHL4, the results revealed 15 SNPs, 3 indels and 1 repeat variation as motif (TG)n(G)n/(AC)n(C)n. Among these variations, the deletion 46delC in exon 1 of CATHL4 was detected in a sample of the Ha Giang breed causing a truncated polypeptide as predicted. The average copy number of CATHL4 of Vietnam indigenous cattle breeds using GADPH as a reference gene showed in range from 2.53 to 2.89 copies.
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Bassam B. J., Caetano-Anolles G., 1993. Silver staining of DNA in polyacrylamide gels. Appl Biochem Biotechnol, 42:
–188.
Bickhart D. M., Hou Y., Schroeder S. G., Alkan C., Cardone M. F., Matukumalli L. K., 2012. Copy number variation of individual cattle genomes using next-generation sequencing. Genome Research, 22: 778–790.
Biswajit B., Mahesh C. P., Satyanagalakshmi K., Meenu C., Purusottam M., Bidhan C. D., Sushil K., Sourav M., Kiran T., Krishna M. P., Tirtha K. D., Sachinandan D., 2015. Diversity, antimicrobial action and structure activity relationship of buffalo cathelicidins. PLoS ONE 10(12): e014474.
Dorin J. R., McHugh B. J., Cox S. L., Davidson D. J., 2015. Mammalian antimicrobial peptides: defensins and cathelicidins. Molecular Medicinal Microbiology, Second Edition, Volume 1: 539–565.
Elsik C. G., Tellam R. L., Worley K. C., 2009. The genome sequence of taurine cattle: a window to ruminant biology and evolution. Science, 324: 522–528.
Gillenwaters E. N., Seabury C. M., Elliott J. S., Womack J. E., 2009. Sequence analysis and polymorphism discovery in 4 members of the bovine cathelicidin gene family. Journal of Heredity. 100(2): 241–245.
Kelly A. L., Mezulis S., Yates C. M., Wass N. M., Sternberg M. J. E., 2015. The Phyre2 web portal for protein modeling, prediction and analysis. Nature Protocol, 10: 845–858.
Kumar P., Henikoff S., Pauline C. Ng., 2009. Predicting the effects of coding non-synonymous variants on protein function using the SIFT algorithm. Nature Protocol, 4(7): 1073−1081.
Lazarev V. N., Govorun V. M., 2010. Antimicrobial Peptides and Their Use in Medicine, Apply Biochemistry and Microbiology, 46(9): 803–814.
Lawrence A. K., Stefans M., Christopher M. Y., Mark N. W. and Michael J. E. S, 2015. The Phyre2 web portal for protein modeling, prediction and analysis. Nature Protocols, 10: 845–858.
Mookherjee N., Brown, K. L., Bowdish D. M., Doria S., Falsafi R., Hokamp K., Roche F. M., Mu R., Doho G. H., Pistolic J., Powers J. P., Bryan J, Brinkman F. S., Hancock R. E., 2006. Modulation of the TLR-mediated inflammatory response by the endogenous human host defense peptide LL-37. Journal of Immunology, 176: 2455–2464.
Ningbo C., Weiwei F., Jianbang Z., Jiafei S., Qiuming C., Zhuqing Z., Hong C., Tad S. S., Chuzhao L., Yu J., 2020. BGVD: An Integrated Database for Bovine Sequencing Variations and Selective Signatures. Genomics Proteomics Bioinformatics, 18: 186–193.
Tomasinsig L., De Conti G., Skerlavaj B., Piccinini R., Mazzilli M., D’Este F., Tossi A., Zanetti M., 2010. Broad-spectrum activity against bacterial mastitis pathogens and activation of mammary epithelial cells support a protective role of neutrophil cathelicidins in bovine mastitis. Infection and Immunity, 78: 1781–1788.
Tomasinsig L., Zanetti M., 2005. The cathelicidins- structure, function and evolution. Current Protein and Peptide Science, 6: 23–34.
Whelehan J. A., Russel N. B., Whelan M. A., 2003. A method for the absolute quantification of cDNA using real-time PCR. Journal of Immunology Methods, 278(1-2): 261–269.
Whelehan C. J., Barry-Reidy A., Meade K. G., Eckersall P. D., Aspinas C., Fernando N., Andrew T. L., Cliona O’F., 2014. Characterisation and expression profile of the bovine cathelicidin gene repertoire in mammary tissue. BMC Genomics, 15: 128.
Zanetti M., 2005. The role of cathelicidins in the innate host defenses of mammals. Current Issues of Molecular Biology, 7: 179–196.
Zanetti M., Del S. G., Storici P., Schneider C., Romeo D., 1993. The cDNA of the neutrophil antibiotic Bac5 predicts a prosequence homologous to a cysteine proteinase inhibitor that is common to other neutrophil antibiotics. The Journal of Biological Chemistry, 268: 522–526.
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Copyright (c) 2023 Thi Tuoi Do, Hong Ngoc Ta, Thi Thuy Anh Tran, Doan Lan Pham, Thi Phuc Do, Thi Hong Van Nguyen
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