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Molecular characterization and expression analysis of osteopontin cDNA from lactating mammary gland in yak (Bos grunniens)

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Abstract

Osteopontin (OPN) is a secreted phosphorylated glycoprotein. It has an important role in mammary gland development and lactation, as well as, is thought to be a potential candidate gene for lactation traits. In the present work, we isolated and characterized a full-length open reading frame (ORF) of yak OPN cDNA from lactating mammary tissue, and examined its expression pattern in mammary gland during different stages of lactation, as well as, the recombinant OPN protein of yak was expressed successfully in E. coli. The sequencing results indicated that the isolated cDNA was 1132-bp in length containing a complete ORF of 837-bp. It encoded a precursor protein of yak OPN consisting of 278 amino acid with a signal peptide of 16 amino acids. Yak OPN has a predicted molecular mass of 29285.975 Da and an isoelectric point of 4.245. It had an identity of 65.50–99.16% in cDNA, identity of 52.06–98.56% and similarity of 65.40–98.56% in deduced amino acids with the corresponding sequences of cattle, buffalo, sheep, goat, pig, human, and rabbit. The phylogenetic analysis indicated that yak OPN had the closest evolutionary relationship with that of cattle, and next buffalo. In mammary gland, yak OPN was generally transcribed in a declining pattern from colostrum period to dry period with an apparent increase of OPN expression being present in the late period of lactation compared with peak period of lactation. Western blot analysis indicated that His-tagged yak OPN protein expressed in E. coli could be recognized not only by an anti-His-tag antibody but also by an anti-human OPN antibody. These results from the present work provided a foundation for further insight into the role of OPN gene in yak lactation.

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References

  1. Zi XD, Lu H, Yin RH, Chen SW (2008) Development of embryos after in vitro fertilization of bovine oocytes with sperm from either yaks (Bos grunniens) or cattle (Bos Taurus). Anim Reprod Sci 108:208–215

    Article  PubMed  Google Scholar 

  2. Lin YQ, Wang GS, Feng J, Huang JQ, Xu YO, Jin SY, Li YP, Jiang ZR, Zheng YC (2011) Comparison of enzyme activities and gene expression profiling between yak and bovine skeletal muscles. Livest Sci 135:93–97

    Article  Google Scholar 

  3. Sasaki M (1994) Yak: Hardy multi-purpose animal of Asiahighland. In: Proceedings of 1st international congress Yak, Lanzhou, China, J Gansu Agric Univ Supplement, pp 1–5

  4. Wu JP, Ma BY, Ren HW, Zhang LP, Xiang Y, Brown MA (2007) Characterization of metallothioneins (MT-I and MT-II) in the yak. J Anim Sci 85:1357–1362

    Article  PubMed  CAS  Google Scholar 

  5. Wang YS, Zou SX, (1995) Protein in milk. In: Biochemistry of milk. Jilin Univ. Press, Changchun, China, pp 1–48

  6. Bai WL, Yin RH, Zheng YC, Ma ZJ, Zhong JC, Yin RL, Dou QL, Zhang SC, Luo GB, Zhao ZH (2010) Cloning and molecular characterization of a yak a-lactalbumin cDNA from mammary tissue. Livest Sci 129:122–128

    Article  Google Scholar 

  7. Nagatomo T, Ohga S, Takada H, Nomura A, Hikino S, Imura M, Ohshima K, Hara T (2004) Microarray analysis of human milk cells: persistent high expression of osteopontin during the lactation period. Clin Exp Immunol 138:47–53

    Article  PubMed  CAS  Google Scholar 

  8. Sodek J, Ganss B, McKee MD (2000) Osteopontin. Crit Rev Oral Biol Med 11:279–303

    Article  PubMed  CAS  Google Scholar 

  9. Schack L, Lange A, Kelsen J, Agnholt J, Christensen B, Petersen TE, Sørensen ES (2009) Considerable variation in the concentration of osteopontin in human milk, bovine milk, and infant formulas. J Dairy Sci 92:5378–5385

    Article  PubMed  CAS  Google Scholar 

  10. Nemir M, Bhattacharyya D, Li X, Singh K, Mukherjee AB, Mukherjee BB (2000) Targeted inhibition of Osteopontin expression in the mammary gland causes abnormal morphogenesis and lactation deficiency. J Biol Chem 275:969–976

    Article  PubMed  CAS  Google Scholar 

  11. Naficy AB, Abu-Elyazeed R, Holmes JL, Rao MR, Savarino SJ, Kim Y, Wierzba TF, Peruski L, Lee YJ, Gentsch JR, Glass RI, Clemens JD (1999) Epidemiology of rotavirus diarrhea in Egyptian children and implications for disease control. Am J Epidemiol 150:770–777

    PubMed  CAS  Google Scholar 

  12. Mastretta E, Longo P, Laccisaglia A, Balbo L, Russo R, Mazzaccara A, Gianino P (2002) Effect of Lactobacillus GG and breast-feeding in the prevention of rotavirus nosocomial infection. J Pediatr Gastroenterol Nutr 35:527–531

    Article  PubMed  Google Scholar 

  13. Bayless KJ, Davis GE, Meininger GA (1997) Isolation and biological properties of osteopontin from bovine milk. Protein Expr Purif 9:309–314

    Article  PubMed  CAS  Google Scholar 

  14. Ron M, Kliger D, Feldmesser E, Seroussi E, Ezra E, Weller JI (2001) Multiple quantitative trait locus analysis of bovine chromosome 6 in the Israeli Holstein population by a daughter design. Genetics 159:727–735

    PubMed  CAS  Google Scholar 

  15. Schnabel RD, Kim JJ, Ashwell MS, Sonstegard TS, Van Tassell CP, Connor EE, Taylor JF (2005) Fine-mapping milk production quantitative trait loci on BTA6: analysis of the bovine osteopontin gene. Proc Natl Acad Sci USA 102:6896–6901

    Google Scholar 

  16. Leonard S, Khatib H, Schutzkus V, Chang YM, Maltecca C (2005) Effects of the osteopontin gene variants on milk production traits in dairy cattle. J Dairy Sci 88:4083–4086

    Article  PubMed  CAS  Google Scholar 

  17. Khatib H, Zaitoun I, Wiebelhaus-Finger J, Chang YM, Rosa GJ (2007) The association of bovine PPARGC1A and OPN genes with milk composition in two independent Holstein cattle populations. J Dairy Sci 90:2966–2970

    Article  PubMed  CAS  Google Scholar 

  18. Oldberg A, Franzen A, Heinegard D (1986) Cloning and sequence analysis of rat bone sialoprotein (osteoprotein) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc Natl Acad Sci USA 83:8819-8823

    Google Scholar 

  19. Craig AM, Smith JH, Denhardt DT (1989) Osteopontin, a transformation-associated cell adhesion phosphoprotein, is induced by 12-o-tetradecanoylphorbol-13-acetate in mouse epidermis. J Biol Chem 264:9682–9689

    PubMed  CAS  Google Scholar 

  20. Wrana JL, Zhang Q, Sodek J (1989) Full length cDNA sequence of porcine secreted phosphoprotein-1 (SPP-I, osteopontin). Nucleic Acids Res 17:10119

    Article  PubMed  CAS  Google Scholar 

  21. Kiefer MC, Bauer DM, Barr PJ (1989) The cDNA and derived amino acid sequence for human osteopontin. Nucleic Acids Res 17:3306

    Article  PubMed  CAS  Google Scholar 

  22. Kerr JM, Fisher LW, Termine JD, Young MF (1991) The cDNA cloning and RNA distribution of bovine osteopontin. Gene 108:237–243

    Article  PubMed  CAS  Google Scholar 

  23. Tantia MS, Mishra B, Bharani Kumar ST, Mishra BP, Kataria RS, Mukesh M, Vijh RK (2008) Characterization of osteopontin gene of Bubalus bubalis. Animal 2:987–990

    Article  CAS  Google Scholar 

  24. Crivello JF, Delvin E (1992) Isolation and characterization of a cDNA for osteopontin-k, a kidney cell adhesion molecule with high homology to osteopontins. J Bone Mine Res 7:693–699

    Article  CAS  Google Scholar 

  25. Koringa PG, Patel AK, Nandasana KN, Ramani UV, Bhong CD, Joshi VK, Wadhvani KN, Kelawala NH, Barvalia DR (2008) Mammary gland biopsy in pregnant and lactating buffaloes in standing position. Indian J Vet Surg 29:50–51

    Google Scholar 

  26. Bai WL, Yin RH, Zhao SJ, Zheng YC, Zhong JC, Zhao ZH (2008) Characterization of a κ-casein genetic variant in the Chinese yak, Bos grunniens. J Dairy Sci 92:1354–1360

    Article  Google Scholar 

  27. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  28. Deléage G, Combet C, Blanchet C, Geourjon C (2001) ANTHEPROT: an integrated protein sequence analysis software with client/server capabilities. Comput Biol Med 31:259–267

    Article  PubMed  Google Scholar 

  29. Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA 4: molecular evolutionary genetics analysis (MEGA) software (version 4.0). Mol Biol Evol 24:1596–1599

    Article  PubMed  CAS  Google Scholar 

  30. Tu ZC, Qiu H, Zhang YP (2002) Polymorphism in mitochondrial DNA (mtDNA) of yak (Bos grunniens). Biochem Genet 30:187–193

    Article  Google Scholar 

  31. Lai SJ, Chen SY, Liu YP, Yao YG (2007) Mitochondrial DNA sequence diversity and origin of Chinese domestic yak. Anim Genet 38:77–80

    Article  PubMed  Google Scholar 

  32. Qi XB, Jianlin H, Wang G, Rege JE, Hanotte O (2010) Assessment of cattle genetic introgression into domestic yak populations using mitochondrial and microsatellite DNA markers. Anim Genet 2010(41):242–252

    Article  Google Scholar 

  33. Bai WL, Yin RH, Dou QL, Jiang WQ, Zhao SJ, Ma ZJ, Luo GB, Zhao ZH (2011) Molecular characterization and phylogenetic analysis of a yak (Bos grunniens) κ-casein cDNA from lactating mammary gland. Mol Biol Rep 38:2711–2718

    Article  PubMed  CAS  Google Scholar 

  34. Sun J, Luo J, Liu JX, Li DQ (2009) Expression of OPN gene during different lactation stages in mammary gland of dairy goat and its effect on growth of MCF-7 cell line. Yi Chuan 31:825–830

    Article  PubMed  CAS  Google Scholar 

  35. Rittling SR, Novick KE (1997) Osteopontin expression in mammary gland development and tumorigenesis. Cell Growth Differ 8:1061–1069

    PubMed  CAS  Google Scholar 

  36. Lampe MA, Patarca A, Iregui MV, Cantor H (1991) Polyclonal B-cell activation by the Eta-1 cytokine and the development of autoimmune disease. J Immunol 147:2902–2906

    PubMed  CAS  Google Scholar 

  37. Ogra PL, Losonsky GA, Fishaut M (1983) Colostrum-derived immunity and maternal-neonatal interaction. Ann NY Acad Sci 409:82–95

    Article  PubMed  CAS  Google Scholar 

  38. Thanh T, Chi VT, Abdullah MP, Omar H, Noroozi M, Napis S (2011) Cloning and characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) cDNA from green microalga Ankistrodesmus convolutus. Mol Biol Rep. doi:10.1007/s11033-011-0679-4

  39. Dando PM, Fortunato M, Strand GB, Smith TS, Barrett AJ (2003) Pyroglutamyl-peptidase I: cloning, sequencing, and characterisation of the recombinant human enzyme. Protein Expr Purif 28:111–119

    Article  PubMed  CAS  Google Scholar 

  40. Kilbane Z, Vaas PR, Cuív PO, O’Connor B (2007) Cloning and heterologous expression of bovine pyroglutamyl peptidase type-1 in Escherichia coli: purification, biochemical and kinetic characterisation. Mol Cell Biochem 297:189–197

    Article  PubMed  CAS  Google Scholar 

  41. Baneyx F (1999) Recombinant protein expression in Escherichia coli. Curr Opin Biotechnol 10:411–421

    Article  PubMed  CAS  Google Scholar 

  42. Meng F, Shen C, He Y, Miao F, Zhang J (2009) Cloning and expression of three peptide-linked β2-microglobulin molecules in Escherichia coli with an isocaudamer technique. Biotechnol Lett 31:831–836

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

We thank the anonymous reviewers for their time and energy in reviewing this article. This work was supported by a grant from Genetically Modified Organisms Breeding Major Projects of China (No.2009ZX08009-156B).

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Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110866, China

    W. L. Bai, R. H. Yin, W. Q. Jiang & G. B. Luo

  2. Research Academy of Animal Husbandry and Veterinary Medicine Science of Jilin Province, Changchun, 130062, China

    R. L. Yin

  3. Institute of Biotechnology, Animal Science Research Academy of Sichuan Province, Chengdu, 610066, China

    S. J. Zhao

  4. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130062, China

    C. Li

  5. College of Animal Science and Veterinary Medicine, Jilin University, Changchun, 130062, China

    R. J. Yang & Z. H. Zhao

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  1. W. L. Bai
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Correspondence to R. H. Yin or Z. H. Zhao.

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W. L. Bai and R. J. Yang contributed equally to this work.

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Bai, W.L., Yang, R.J., Yin, R.H. et al. Molecular characterization and expression analysis of osteopontin cDNA from lactating mammary gland in yak (Bos grunniens). Mol Biol Rep 39, 3627–3635 (2012). https://doi.org/10.1007/s11033-011-1137-z

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  • DOI: https://doi.org/10.1007/s11033-011-1137-z

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