Characterization of a novel antimicrobial peptide gene from the reproductive tract of indigenous cows (Bos indicus) of Asom

REZINA SULTANA; D J KALITA; S SARMA; A BARUAH; B DEVI

doi:10.56093/ijans.v86i2.55778

Authors

REZINA SULTANA Assam Agricultural University, Khanapara, Guwahati, Asom 781 022 India
D J KALITA Assam Agricultural University, Khanapara, Guwahati, Asom 781 022 India
S SARMA Assam Agricultural University, Khanapara, Guwahati, Asom 781 022 India
A BARUAH Assam Agricultural University, Khanapara, Guwahati, Asom 781 022 India
B DEVI Assam Agricultural University, Khanapara, Guwahati, Asom 781 022 India

https://doi.org/10.56093/ijans.v86i2.55778

Keywords:

Bos indicus, cDNA, Female reproductive tract, Lingual antimicrobial peptide, RNA

Abstract

Antimicrobial peptides are innate immune defense peptides protecting against infection. Defensins and cathelicidins are the two major antimicrobial peptides in eukaryotes. In the present study, female reproductive tract was collected from apparently healthy local cows (Bos indicus) of Asom after slaughter. cDNA was synthesized from the extracted RNA by reverse transcription and amplified the Lingual Antimicrobial Peptide (LAP) gene (227 bp) using specific primers. The purified product was sequenced and sequence were aligned Nucleotide sequence was BLAST with twelve published sequences and analyzed using DNA Star software. At nucleotide level, Bos indicus LAP of reproductive tract showed the highest similarity of 97.4% with Bos taurus LAP of tongue followed by 92.8% with buffalo EBD. We identified the highest similarity (93.8%) of Bos indicus LAP of reproductive tract with Bos taurus LAP of tongue followed by buffalo EBD (86.2%). The phylogenetic analyses at nucleotide and amino acid level showed that Bos indicus LAP of reproductive tract and Bos taurus LAP of tongue are closely evolutionarily which implied that they might have diverged from ancestral gene. We conclude that female reproductive-tract epithelium of local cows of Asom express a potent AMP similar to that of Bos taurus LAP of tongue.

Downloads

Download data is not yet available.

References

Bals R. 2000. Epithelial antibacterial peptides in host defense against infection. Respiratory Research 1: 141–50. DOI: https://doi.org/10.1186/rr25

Bishop S C and Woolliams J A. 2014. Genomics and disease resistance studies in livestock. Livestock Science 166: 190– 98. DOI: https://doi.org/10.1016/j.livsci.2014.04.034

Boman H G. 1995.Peptide antibiotics and their role in innate immunity. Annual Review of Immunology 13 :61–92. DOI: https://doi.org/10.1146/annurev.iy.13.040195.000425

Das D K, Sharma B, Mitra A and Kumar A. 2005. Molecular cloning and characterization of ß-defensin cDNA expressed in distal ileum of buffalo (Bubalus bubalis). DNA Seqence 16: 16–20. DOI: https://doi.org/10.1080/10425170400020399

Diamond G, Zasloff M, Eck H, Brasseur M, Maloy W L and Bevins C L. 1991. Tracheal antimicrobial peptide, a cysteine- rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. Proceedings of National Academy of Science 88:3952–56. DOI: https://doi.org/10.1073/pnas.88.9.3952

Fearon D and Locksley R. 1996. The instructive role of innate immunity in the acquired immune response. Science 272: 50– 54. DOI: https://doi.org/10.1126/science.272.5258.50

Garica J R, Krause A and Scchulz S. 2001. Fimpong-Boateng A. Human beta defensin 4: a novel inducible peptide with a specific salt-sensitive spectrum of antimicrobial activity. Federation of American Societies for Experimental Biology journal 15: 1819–21. DOI: https://doi.org/10.1096/fj.00-0865fje

Ganz T and Lehrer R I. 1994. Defensins. Current Opinion in Immunology 6: 584–89. DOI: https://doi.org/10.1016/0952-7915(94)90145-7

Gennaro R, Zanetti M, Benincasa M, Podda E and Miani M. 2002. Pro-rich antimicrobial peptides from animals: structure, biological functions and mechanism of action. Current Pharmacological Design 8: 763–78. DOI: https://doi.org/10.2174/1381612023395394

Grandjean V, Vincent S, Martin L, Rassoulzadegan M and Cuzin F. 1997. Antimicrobial protection of the mouse testis : Synthesis of defensin of the cryptdin family. Biology of Reproduction 57: 1115–22. DOI: https://doi.org/10.1095/biolreprod57.5.1115

Gorbach S L. 2001.Antimicrobial use in animal feed: time to stop. New England Journal of Medicine 345: 1202–03. DOI: https://doi.org/10.1056/NEJM200110183451610

Joseph D and More T. 2011. Molecular characterization of Lingual antimicrobial peptide in the female reproductive tract to buffalo. Veterinary World 4: 120–23. DOI: https://doi.org/10.5455/vetworld.2011.120-123

Kalita D J and Kumar A. 2009. Molecular cloning and characterization of lingual antimicrobial peptide cDNA of Bubalus bubalis. Veterinary Science 86:91–97. DOI: https://doi.org/10.1016/j.rvsc.2008.05.002

Li P, Bin C C, Chung H H, Chung S S, Shang Y W and Zhang Q Y. 2001. An antimicrobial peptide gene found in the male reproductive system of rats. Science 291: 1783–85. DOI: https://doi.org/10.1126/science.1056545

Malm J, Sorenson O, Persson T, Frohm-Nilsson M, Johansson B, Bjartell A, et al. 2000. The human cationic antimicrobial protein (hCAP18) is expressed in the epithelium of human epididymis, is present in seminal plasma at high concentration and is attached to spermatozoa. Infection and Immunity 68: 4297–302. DOI: https://doi.org/10.1128/IAI.68.7.4297-4302.2000

Markus Reinholz M D, Thomas Ruzicka M D and Jürgen Schauber M D. 2012. Cathelicidin LL–37: An Antimicrobial Peptide with a Role in Inflammatory Skin Disease. Annales de Dermatologie et de Vénéréologie 24: 126–35. DOI: https://doi.org/10.5021/ad.2012.24.2.126

Park S, Kim H, Kim C, Yun H, Choi E and Lee B. 2002. Role of proline, cysteine and disulphide bridge in the structure and activity of the antimicrobial peptide gaegurin5. Biochemical Journal 368: 171–82. DOI: https://doi.org/10.1042/bj20020385

Ravi C, Jeyashree A R and Devi K. 2011. Antimicrobial Peptides from Insects: An Overview. Research in Biotechnology 2: 1– 7.

Robert E W, Hancock P and Hans-Georg S. 2006. Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nature Biotechnology 24: 1551–57. DOI: https://doi.org/10.1038/nbt1267

Schonwetter BS, Stolzenberg ED and Zasloff MA. 1995. Epithelial antibiotic induced at sites of inflammation. Science 267: 1645– 48. DOI: https://doi.org/10.1126/science.7886453

Sharma A, Kumar Ashish, Kumar A and Dev K. 2010. Characterization of goat lingual antimicrobial peptide cDNA. Journal of Immunology and Immunopathology 12: 46–51.

Stolzenberg E D, Anderson G M, Ackermann M R, Whitlock R H and Zasloff M. 1997. Epithelial antibiotic induced in states of disease. Proceedings of National Academy of Science 94: 8686–90. DOI: https://doi.org/10.1073/pnas.94.16.8686

Suh J Y, Lee Y T, Park C B, Lee K H, Kim S C and Choi B S. 1999. Structural and functional implications of a proline residue in the antimicrobial peptide gaegurin. European Jornal of Biochemistry 266: 665–74. DOI: https://doi.org/10.1046/j.1432-1327.1999.00917.x

Tarver A P, Clark D P, Diamond G, Russell J P, Erdjument- Bromage H, Tempst P, Cohen K S, Jones D E, Sweeney R W, Wines M, Hwang S and Bevins C L. 1998. Enteric beta- defensin: molecular cloning and characterization of a gene with inducible intestinal epithelial cell expression associated with Cryptosporidium parvum infection. Infection and Immunology 66: 1045–56. DOI: https://doi.org/10.1128/IAI.66.3.1045-1056.1998

Yang Y, Jing L, Li T, Cao G and Liu S. 2011. A new Beta defensin from sika deer: molecular cloning and sequence characterization. Animal Biotechnology 22: 64–70. DOI: https://doi.org/10.1080/10495398.2011.554104