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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 9 (september 2019) : 1129-1134,   Doi: 10.18805/ijar.B-3636

Molecular cloning and expression kinetics of serum interferon stimulated gene for early pregnancy detection

K
Kanisht Batra
T
Trilok Nanda
A
Aman Kumar
V
Vinay kumar
G
Gopal Jee Gopal
S
Sushila Maan
1Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125 004, Haryana, India.
Cite article:- Batra Kanisht, Nanda Trilok, Kumar Aman, kumar Vinay, Gopal Jee Gopal, Maan Sushila (2018). Molecular cloning and expression kinetics of serum interferon stimulated gene for early pregnancy detection. Indian Journal of Animal Research. 53(9): 1129-1134. doi: 10.18805/ijar.B-3636.

ABSTRACT

Interferon stimulated protein 15 is a well-known ubiquitin cross-reactive protein which is released from the ruminant uterus in response to interferon-tau (IFN-t) during conceptus implantation. This protein increases significantly during the early stages of pregnancy. Therefore, in the present study, Interferon stimulated gene (ISG15) coding region was isolated from buffalo blood at day 18 of pregnancy after artificial insemination. ISG15 was cloned in pET100 vector, overexpressed in E.coli and purified to investigate its properties. The protein was immunoblotted with mouse anti-His antibody to evaluate its characteristics after purification. To conclude, elucidation of protein properties in the prokaryotic system will provide an opportunity for understanding the basic biology of pre and post-implantation and development of a tool for early pregnancy diagnosis. 

REFERENCES

  1. Abdullah, M., Mohanty, T.K., Patbandha, T.K., Bhakat, M., Madkar, A.R., Kumaresan A. and Mohanty, A.K. (2017). Pregnancy diagnosis-positive rate and conception rate as indicator of farm reproductive performance. Indian J. Anim. Res., 51 (1): 170-174.
  2. Austin, K.J., Ward, S.K., Teixeira, M.G., Dean, V.C., Moore, D.W. and Hansen, T.R. (1996). Ubiquitin cross-reactive protein is released by the bovine uterus in response to interferon during early pregnancy. Biol. Reprod.,54: 600-606.
  3. Austin, K.J., Bany, B.M., Belden, E.L., Rempel, L.A., Cross, J.C. and Hansen, T.R. (2003). Interferon-stimulated gene-15 (Isg15) expression is up-regulated in the mouse uterus in response to the implanting conceptus. Endocrinology, 144: 3107-3113.
  4. Austin, K.J., Carr, A.L., Pru, J.K., Hearn, C.E., George, E.L., Belden, E.L. and Hansen, T.R. (2004). Localization of ISG15 and conjugated proteins in bovine endometrium using immunohistochemistry and electron microscopy. Endocrinology, 145: 967-975.
  5. Batra, K., Nanda, T., Kumar, A., Gupta, A.K., Kumari, R., Kumar, V., Sheoran, N. and Maan, S., (2018). Molecular cloning and characterization of Mx2 for early pregnancy diagnosis in Bubalus bubalis. Proc. Natl. Acad. Sci. India, Sect. B Biol. Sci. https://doi.org/10.1007/s40011-018-0993-x
  6. Batra, K. (2017). In vitro expression, molecular and immunological characterization of early pregnancy-related interferon tau stimulated gene of Bubalus bubalis. Ph.D Dissertation, LUVAS, Hisar, Haryana, India.
  7. Bebington, C., Doherty, F.J. and Fleming, S.D. (1999). Ubiquitin cross-reactive protein gene expression is increased in decidualized endometrial stromal cells at the initiation of pregnancy. Mol. Hum. Reprod.,5: 966-972.
  8. Bhardwaj A., Kumar, S., Nayan, Varij., Sharma, P., Pal, Y. and Yadav, S.C. (2018). Expression and characterization of recombinant single chain beta-alpha equine chorionic gonadotropin in prokaryotic host. Indian J. Anim. Res., DOI: 10.18805/ijar.B-3571.
  9. Bott, R.C., Ashley, R.L., Henkes, L.E., Antoniazzi, A.Q., Bruemmer, J.E., Niswender, G.D., Bazer, F.W., Spencer, T.E., Smirnova, N.P., Anthony, R.V. and Hansen, T.R. (2010). Uterine vein infusion interferon tau (IFNT) extends luteal life span in ewes. Biol. Reprod.,82: 725-735.
  10. Cicek, H., Tandogan, M. and Uyarlar, C. (2017). Financial losses due to fertility problems in Anatolian Dairy Buffalo. Indian J. Anim. Res., 51 (6): 1144-1148.
  11. Chen, Y., Green, J.A., Antoniou, E., Ealy, A.D., Mathialagan, N., Walker, A.M., Avalle, M.P., et al. (2006). Effect of interferon-tau administration on endometrium of nonpregnant ewes: a comparison with pregnant ewes. Endocrinology, 147: 2127-2137.
  12. Debeljak, N., Feldman, L., Davis, K. L., Komel, R. and Sytkowski, A. J. (2006). Variability in the Immunodetection of His-tagged Recombinant Proteins. Anal. Biochem., 359(2): 216-223.
  13. Farin, C.E., Imakawa, K., Hansen, T.R, McDonnell, J.J., Murphy, C.N., Farin, P.W. and Roberts, R.M. (1990). Expression of trophoblastic interferon genes in sheep and cattle. Biol. Reprod., 43: 210-218.
  14. Gifford, C.A., Racicot, K., Clark, D.S., Austin, K.J., Hansen, T.R., Lucy, M.C., Davies, C.J. and Ott, T.L. (2007). Regulation of interferon-    stimulated genes in peripheral blood leukocytes in pregnant and bred, nonpregnant dairy cows. J. Dairy Sci., 90: 274-280.
  15. Green, J.C., Okamura, C.S., Poock, S.E. and Lucy, M.C. (2010). Measurement of interferon-tau (IFN-tau) stimulated gene expression in blood leukocytes for pregnancy diagnosis within 18-20d after insemination in dairy cattle. Anim. Reprod. Sci., 121: 24-33.
  16. Haq, I.U., Han, Y., Ali, T., Wang, Y., Gao, H., Lin, L., Wu, Y., Wu, S. and Zeng, S. (2016). Expression of interferon-stimulated gene ISG15 and ubiquitination enzymes in peripheral blood monocyte during early pregnancy in dairy cattle. Reprod. Biol., 16(4): 255-260.
  17. Han, H., Austin, K.J., Rempel, L.A. and Hansen, T.R. (2006). Low blood ISG15 mRNA and progesterone levels are predictive of non-    dairy cows. J. Endocrinol., 191: 505-512.
  18. Hansen, T.R., Austin, K.J. and Johnson, G.A. (1997). Transient ubiquitin cross-reactive protein gene expression in the bovine pregnant endometrium. Endocrinology, 138: 5079-5082.
  19. Hansen, T.R., Austin, K.J., Perry, D.J., Pru, J.K., Teixeira, M.G. and Johnson, G.A. (1999). Mechanism of action of interferon-t in the uterus during early pregnancy. J. Reprod. Fertil. Suppl.,54: 329-339.
  20. Jain, A., Baviskar, P.S., Kandasamy, S., Kumar, R., Singh, R., Kumar, S., Agarwal, S.K., Joshi, P. and Mitra, A. (2012). Interferon stimulated gene 15 (ISG15): molecular characterization and expression profile in endometrium of buffalo (Bubalus bubalis). Anim. Reprod. Sci., 133(3-4):159-168.
  21. Jamuna, V., Chakravarty, A.K., Singh, A. and Patil, C.S. (2015). Genetic parameters of fertility and production traits in Murrah buffaloes.    Indian J. Anim. Res., 49 (3): 288-291.
  22. Johnson, G.A., Austin, K.J., Collins, A.M., Murdoch, W.J. and Hansen, T.R. (1999). Endometrial ISG17 mRNA and a related mRNA are induced by interferon-t and localized to glandular epithelial and stromal cells from pregnant cows. Endocrine, 10: 243-252.
  23. Joyce, M.M., White, F.J., Burghardt, R.C., Muniz, J.J., Spencer, T.E., Bazer, F.W. and Johnson, G.A. (2005). Interferon stimulated gene 15 conjugates to endometrial cytosolic proteins and is expressed at the uterine-placental interface throughout pregnancy in sheep. Endocrinology, 146: 675-684.
  24. Kang, D., Gho, Y., Suh, M.K. and Kang, C.H. (2002). Highly sensitive and fast protein detection with Coomassie brilliant blue in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Bull. Korean. Chem. Soc., 23(11): 1511-1512.
  25. Kim, M.S., Min, K.S. and Imakawa, K. (2013). Regulation of interferon-stimulated gene (ISG) 12, ISG15, and MX1 and MX2 by conceptus interferons (IFNTs) in bovine uterine epithelial Cells. Asian-Australas J. Anim. Sci., 26(6): 795-803.
  26. Nallamsetty, S. and Waugh, D.S. (2007). A generic protocol for the expression and purification of recombinant proteins in Escherichia coli using a combinatorial His6-maltose binding protein fusion tag. Nat. Protoc., 2(2): 383-391.
  27. Oliveira, J.F., Henkes, L.E., Ashley, R.L., Purcell, S.H., Smirnova, N.P., Veeramachaneni, D.N., Anthony, R.V. and Hansen, T.R. (2008). Expression of interferon (IFN)-stimulated genes in extrauterine tissues during early pregnancy in sheep is the consequence of endocrine IFN-tau release from the uterine vein. Endocrinology, 149: 1252-1259.
  28. Perry, D.J., Austin, K.J. and Hansen, T.R. (1999). Cloning of IFN-stimulated gene 17: the promoter and nuclear proteins that regulates transcription. Mol. Endocrinol., 13: 1197-1206.
  29. Roberts, R.M., Klemann, S.W., Leaman, D.W., Bixby, J.A., Cross, J.C., Farin, C.E., Imakawa, K. and Hansen, T.R. (1991). The polypeptides and genes for ovine and bovine trophoblast protein-1. J. Reprod. Fertil. Suppl.,43: 3-12.
  30. Rosano, G.L. and Ceccarelli, E.A. (2014). Recombinant protein expression in Escherichia coli: advances and challenges. Front. Microbiol., 5: 172. 
  31. Soumya, N., Das, D., Jeyakumar, S., Mondal, S., Mor, A. and Mundhe, U. (2016). Differential expression of ISG 15 mRNA in peripheral blood mononuclear cells of nulliparous and multiparous pregnant versus non-pregnant Bos indicus cattle. Reprod. Domest. Anim., 52: 97-106.
  32. Sulkowski, E. (1987). In Protein Purification: Micro to Macro, Burgess ed.R. (Liss, New York), pp. 149-162.
  33. Thatcher, W.W., Meyer, M.D. and Danet-Desnoyers, G. (1995). Maternal recognition of pregnancy. J. Reprod. Fertil. Suppl.,49: 15-28.
  34. Yang, L., Wang, X.L., Wan, P.C., Zhang, L.Y., Wu, Y., Tang, D.W. and Zeng, S.M. (2010). Up-regulation of expression of interferon-    stimulated gene 15 in the bovine corpus luteum during early pregnancy. J. Dairy Sci., 93: 1000-1011.
  35. Yang, L., Wang, Y., Xingshu, M.A., Wang, S. and Zhang, L. (2016). Changes in expression of Th1 and Th2 cytokines in bovine peripheral blood mononuclear cells during early pregnancy. Indian J. Anim. Res., 50 (4): 466-470.
  36. Zerbs, S., Giuliani, S. and Collart, F. (2014). Small-scale expression of proteins in E. coli. Methods Enzymol., 536:117-31. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 9 (september 2019) : 1129-1134,   Doi: 10.18805/ijar.B-3636

    Molecular cloning and expression kinetics of serum interferon stimulated gene for early pregnancy detection

    K
    Kanisht Batra
    T
    Trilok Nanda
    A
    Aman Kumar
    V
    Vinay kumar
    G
    Gopal Jee Gopal
    S
    Sushila Maan
    1Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar-125 004, Haryana, India.
    Cite article:- Batra Kanisht, Nanda Trilok, Kumar Aman, kumar Vinay, Gopal Jee Gopal, Maan Sushila (2018). Molecular cloning and expression kinetics of serum interferon stimulated gene for early pregnancy detection. Indian Journal of Animal Research. 53(9): 1129-1134. doi: 10.18805/ijar.B-3636.

    ABSTRACT

    Interferon stimulated protein 15 is a well-known ubiquitin cross-reactive protein which is released from the ruminant uterus in response to interferon-tau (IFN-t) during conceptus implantation. This protein increases significantly during the early stages of pregnancy. Therefore, in the present study, Interferon stimulated gene (ISG15) coding region was isolated from buffalo blood at day 18 of pregnancy after artificial insemination. ISG15 was cloned in pET100 vector, overexpressed in E.coli and purified to investigate its properties. The protein was immunoblotted with mouse anti-His antibody to evaluate its characteristics after purification. To conclude, elucidation of protein properties in the prokaryotic system will provide an opportunity for understanding the basic biology of pre and post-implantation and development of a tool for early pregnancy diagnosis. 

    REFERENCES

    1. Abdullah, M., Mohanty, T.K., Patbandha, T.K., Bhakat, M., Madkar, A.R., Kumaresan A. and Mohanty, A.K. (2017). Pregnancy diagnosis-positive rate and conception rate as indicator of farm reproductive performance. Indian J. Anim. Res., 51 (1): 170-174.
    2. Austin, K.J., Ward, S.K., Teixeira, M.G., Dean, V.C., Moore, D.W. and Hansen, T.R. (1996). Ubiquitin cross-reactive protein is released by the bovine uterus in response to interferon during early pregnancy. Biol. Reprod.,54: 600-606.
    3. Austin, K.J., Bany, B.M., Belden, E.L., Rempel, L.A., Cross, J.C. and Hansen, T.R. (2003). Interferon-stimulated gene-15 (Isg15) expression is up-regulated in the mouse uterus in response to the implanting conceptus. Endocrinology, 144: 3107-3113.
    4. Austin, K.J., Carr, A.L., Pru, J.K., Hearn, C.E., George, E.L., Belden, E.L. and Hansen, T.R. (2004). Localization of ISG15 and conjugated proteins in bovine endometrium using immunohistochemistry and electron microscopy. Endocrinology, 145: 967-975.
    5. Batra, K., Nanda, T., Kumar, A., Gupta, A.K., Kumari, R., Kumar, V., Sheoran, N. and Maan, S., (2018). Molecular cloning and characterization of Mx2 for early pregnancy diagnosis in Bubalus bubalis. Proc. Natl. Acad. Sci. India, Sect. B Biol. Sci. https://doi.org/10.1007/s40011-018-0993-x
    6. Batra, K. (2017). In vitro expression, molecular and immunological characterization of early pregnancy-related interferon tau stimulated gene of Bubalus bubalis. Ph.D Dissertation, LUVAS, Hisar, Haryana, India.
    7. Bebington, C., Doherty, F.J. and Fleming, S.D. (1999). Ubiquitin cross-reactive protein gene expression is increased in decidualized endometrial stromal cells at the initiation of pregnancy. Mol. Hum. Reprod.,5: 966-972.
    8. Bhardwaj A., Kumar, S., Nayan, Varij., Sharma, P., Pal, Y. and Yadav, S.C. (2018). Expression and characterization of recombinant single chain beta-alpha equine chorionic gonadotropin in prokaryotic host. Indian J. Anim. Res., DOI: 10.18805/ijar.B-3571.
    9. Bott, R.C., Ashley, R.L., Henkes, L.E., Antoniazzi, A.Q., Bruemmer, J.E., Niswender, G.D., Bazer, F.W., Spencer, T.E., Smirnova, N.P., Anthony, R.V. and Hansen, T.R. (2010). Uterine vein infusion interferon tau (IFNT) extends luteal life span in ewes. Biol. Reprod.,82: 725-735.
    10. Cicek, H., Tandogan, M. and Uyarlar, C. (2017). Financial losses due to fertility problems in Anatolian Dairy Buffalo. Indian J. Anim. Res., 51 (6): 1144-1148.
    11. Chen, Y., Green, J.A., Antoniou, E., Ealy, A.D., Mathialagan, N., Walker, A.M., Avalle, M.P., et al. (2006). Effect of interferon-tau administration on endometrium of nonpregnant ewes: a comparison with pregnant ewes. Endocrinology, 147: 2127-2137.
    12. Debeljak, N., Feldman, L., Davis, K. L., Komel, R. and Sytkowski, A. J. (2006). Variability in the Immunodetection of His-tagged Recombinant Proteins. Anal. Biochem., 359(2): 216-223.
    13. Farin, C.E., Imakawa, K., Hansen, T.R, McDonnell, J.J., Murphy, C.N., Farin, P.W. and Roberts, R.M. (1990). Expression of trophoblastic interferon genes in sheep and cattle. Biol. Reprod., 43: 210-218.
    14. Gifford, C.A., Racicot, K., Clark, D.S., Austin, K.J., Hansen, T.R., Lucy, M.C., Davies, C.J. and Ott, T.L. (2007). Regulation of interferon-    stimulated genes in peripheral blood leukocytes in pregnant and bred, nonpregnant dairy cows. J. Dairy Sci., 90: 274-280.
    15. Green, J.C., Okamura, C.S., Poock, S.E. and Lucy, M.C. (2010). Measurement of interferon-tau (IFN-tau) stimulated gene expression in blood leukocytes for pregnancy diagnosis within 18-20d after insemination in dairy cattle. Anim. Reprod. Sci., 121: 24-33.
    16. Haq, I.U., Han, Y., Ali, T., Wang, Y., Gao, H., Lin, L., Wu, Y., Wu, S. and Zeng, S. (2016). Expression of interferon-stimulated gene ISG15 and ubiquitination enzymes in peripheral blood monocyte during early pregnancy in dairy cattle. Reprod. Biol., 16(4): 255-260.
    17. Han, H., Austin, K.J., Rempel, L.A. and Hansen, T.R. (2006). Low blood ISG15 mRNA and progesterone levels are predictive of non-    dairy cows. J. Endocrinol., 191: 505-512.
    18. Hansen, T.R., Austin, K.J. and Johnson, G.A. (1997). Transient ubiquitin cross-reactive protein gene expression in the bovine pregnant endometrium. Endocrinology, 138: 5079-5082.
    19. Hansen, T.R., Austin, K.J., Perry, D.J., Pru, J.K., Teixeira, M.G. and Johnson, G.A. (1999). Mechanism of action of interferon-t in the uterus during early pregnancy. J. Reprod. Fertil. Suppl.,54: 329-339.
    20. Jain, A., Baviskar, P.S., Kandasamy, S., Kumar, R., Singh, R., Kumar, S., Agarwal, S.K., Joshi, P. and Mitra, A. (2012). Interferon stimulated gene 15 (ISG15): molecular characterization and expression profile in endometrium of buffalo (Bubalus bubalis). Anim. Reprod. Sci., 133(3-4):159-168.
    21. Jamuna, V., Chakravarty, A.K., Singh, A. and Patil, C.S. (2015). Genetic parameters of fertility and production traits in Murrah buffaloes.    Indian J. Anim. Res., 49 (3): 288-291.
    22. Johnson, G.A., Austin, K.J., Collins, A.M., Murdoch, W.J. and Hansen, T.R. (1999). Endometrial ISG17 mRNA and a related mRNA are induced by interferon-t and localized to glandular epithelial and stromal cells from pregnant cows. Endocrine, 10: 243-252.
    23. Joyce, M.M., White, F.J., Burghardt, R.C., Muniz, J.J., Spencer, T.E., Bazer, F.W. and Johnson, G.A. (2005). Interferon stimulated gene 15 conjugates to endometrial cytosolic proteins and is expressed at the uterine-placental interface throughout pregnancy in sheep. Endocrinology, 146: 675-684.
    24. Kang, D., Gho, Y., Suh, M.K. and Kang, C.H. (2002). Highly sensitive and fast protein detection with Coomassie brilliant blue in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Bull. Korean. Chem. Soc., 23(11): 1511-1512.
    25. Kim, M.S., Min, K.S. and Imakawa, K. (2013). Regulation of interferon-stimulated gene (ISG) 12, ISG15, and MX1 and MX2 by conceptus interferons (IFNTs) in bovine uterine epithelial Cells. Asian-Australas J. Anim. Sci., 26(6): 795-803.
    26. Nallamsetty, S. and Waugh, D.S. (2007). A generic protocol for the expression and purification of recombinant proteins in Escherichia coli using a combinatorial His6-maltose binding protein fusion tag. Nat. Protoc., 2(2): 383-391.
    27. Oliveira, J.F., Henkes, L.E., Ashley, R.L., Purcell, S.H., Smirnova, N.P., Veeramachaneni, D.N., Anthony, R.V. and Hansen, T.R. (2008). Expression of interferon (IFN)-stimulated genes in extrauterine tissues during early pregnancy in sheep is the consequence of endocrine IFN-tau release from the uterine vein. Endocrinology, 149: 1252-1259.
    28. Perry, D.J., Austin, K.J. and Hansen, T.R. (1999). Cloning of IFN-stimulated gene 17: the promoter and nuclear proteins that regulates transcription. Mol. Endocrinol., 13: 1197-1206.
    29. Roberts, R.M., Klemann, S.W., Leaman, D.W., Bixby, J.A., Cross, J.C., Farin, C.E., Imakawa, K. and Hansen, T.R. (1991). The polypeptides and genes for ovine and bovine trophoblast protein-1. J. Reprod. Fertil. Suppl.,43: 3-12.
    30. Rosano, G.L. and Ceccarelli, E.A. (2014). Recombinant protein expression in Escherichia coli: advances and challenges. Front. Microbiol., 5: 172. 
    31. Soumya, N., Das, D., Jeyakumar, S., Mondal, S., Mor, A. and Mundhe, U. (2016). Differential expression of ISG 15 mRNA in peripheral blood mononuclear cells of nulliparous and multiparous pregnant versus non-pregnant Bos indicus cattle. Reprod. Domest. Anim., 52: 97-106.
    32. Sulkowski, E. (1987). In Protein Purification: Micro to Macro, Burgess ed.R. (Liss, New York), pp. 149-162.
    33. Thatcher, W.W., Meyer, M.D. and Danet-Desnoyers, G. (1995). Maternal recognition of pregnancy. J. Reprod. Fertil. Suppl.,49: 15-28.
    34. Yang, L., Wang, X.L., Wan, P.C., Zhang, L.Y., Wu, Y., Tang, D.W. and Zeng, S.M. (2010). Up-regulation of expression of interferon-    stimulated gene 15 in the bovine corpus luteum during early pregnancy. J. Dairy Sci., 93: 1000-1011.
    35. Yang, L., Wang, Y., Xingshu, M.A., Wang, S. and Zhang, L. (2016). Changes in expression of Th1 and Th2 cytokines in bovine peripheral blood mononuclear cells during early pregnancy. Indian J. Anim. Res., 50 (4): 466-470.
    36. Zerbs, S., Giuliani, S. and Collart, F. (2014). Small-scale expression of proteins in E. coli. Methods Enzymol., 536:117-31. 
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