Abstract
A small cDNA fragment containing a ribosome-inactivating site was isolated from the leaf cDNA population of Celosia cristata by polymerase chain reaction (PCR). PCR was conducted linearly using a degenerate primer designed from the partially conserved peptide of ribosome-inactivating/antiviral proteins. Sequence analysis showed that it is 150 bp in length. The cDNA fragment was then cloned in a bacterial expression vector and expressed in Escherichia coli as a ∼57 kD fused protein, and its presence was further confirmed by Western blot analysis. The recombinant protein was purified by affinity chromatography. The purified product showed strong antiviral activity towards tobacco mosaic virus on host plant leaves, Nicotiana glutinosa, indicating the presence of a putative antiviral determinant in the isolated cDNA product. It is speculated that antiviral site is at, or is separate but very close to, the ribosome inactivating site. We nominate this short cDNA frag ment reported here as a good candidate to investigate further the location of the antiviral determinants. The isolated cDNA sequence was submitted to EMBL databases under accession number of AJ535714.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
REFERENCES
Verma, H. N., and Baranwal, V. K. (1995) in Antiviral Proteins from Higher Plants (Chessin, M. B. D., and Zipf, eds.) CRC Press, Boca Raton, pp. 1–21.
Kumar, D., Hridya, N., Verma, N. T., and Krishna, K. T. (1997) Plant Mol. Biol., 33, 745–751.
Kwon, S., An, C. S., Liu, J. R., and Peak, K. (1997) Biosci. Biotech. Biochem., 61, 1613–1614.
Balasaraswathi, R., Sadasivam, S., Ward, M., and Walker, J. M. (1998) Phytochem., 47, 561–565.
Den Hartog, M. T., Lubelli, C., Boon, L., Heerkens, S., Ortiz Buijsse, A. P., de Boer, M., and Stirpe, F. (2002) Eur. J. Biochem., 269, 1772–1779.
Lodge, K., Kaniewaski, W. K., and Tumer, N. E. (1993) Proc. Natl. Acad. Sci. USA, 90, 7089–7093.
Hong, Y. G., Saunders, K., Martin, R., Hartley, and Standly, J. (1996) Virology, 220, 119–127.
Smirnov, S., Shulaev, V., and Tumer, N. E. (1997) Plant Physiol., 114, 1113–1121.
Wang, P., Zoubenko, O., and Tumer, N. E. (1998) Plant Mol. Biol., 38, 957–964.
Guo, Y. J., Demin, J., Man Li, W., Guo, B., Bin, W., Jin, D. M., Weng, M. L., Guo, B. T., and Wang, B. (1999) Acta. Botanica Sinica, 41, 334–336.
Tumer, N. E., Hudak, R. D. R., Coetzer, C., Wang, P., and Zoubenko, O. (1999) in Current Topics in Microbiology and Immunology, Plant Biotechnology: New Products and Applications (Hammond, J., McGarvey, P., and Yusobov, V., eds.) Vol. 240, Springer-Verlag, Berlin Heidelberg, pp. 139–158.
Chen, Y., Vandenbussche, F., Rouge, P., Proost, P., Peumans, W. J., and van Damme, E. J. M. (2002) Eur. J. Biochem., 269, 2897–2906.
Vandenbussche, F., Desmyter, S., Ciani, M., Proost, P., and van Damme, E. J. M. (2004) Eur. J. Biochem., 271, 1508–1515.
Tumer, N. E., Hwang, D. J., and Bonness, M. (1997) Proc. Natl. Acad. Sci. USA, 94, 3866–3871.
Zoubenko, O., Uckun, F., Hur, Y., Chet, I., and Tumer, N. E. (1997) Nature Biotech., 15, 992–996.
Zoubenko, O., Hudak, K., and Tumer, N. E. (2000) Plant Mol. Biol., 44, 219–229.
Hudak, K. A., di Parikh, B. A. R., Baricevic, M., Santana, M., Seskar, M., and Tumer, N. E. (2004) Nucleic Acids Res., 32, 4244–4256.
Balasubramanyam, A., Branwal, V. K., Lodha, M. L., Verma, A., and Kapoor, H. C. (2000) Plant Sci., 154, 13–21.
Baranwal, V. K., Nilgum, E. T., and Kapoor, H. C. (2002) Indian J. Exp. Biol., 40, 1195–1197.
Birnboim, H. C., and Dolly, J. (1979) Nucleic Acids Res., 7, 1513–1515.
Laemmli, U. K. (1970) Nature, 227, 680–685.
O'Donnel, I. J., Shukla, D. D., and Gough, K. H. (1982) J. Virol. Meth., 4, 19–26.
Xu, J., Kaloyanova, D., Ivanov, I. G., and AbouHaidar, M. G. (1998) Arch. Biochem. Biophys., 351, 82–88.
Stripe, F., Barbieri, L., Batelli, M., and Lappi, D. A. (1992) Biotech., 10, 405–412.
Kwon, S. Y., Chung, S. A., Liu, J. R., Kwak, S. S., Lee, H. S., Kim, J. K., and Paek, K. H. (2000) Mol. Cells, 10, 8–12.
Endo, Y., and Tsurugi, K. (1988) J. Biol. Chem., 263, 8735–8739.
Kataoka, J., Habuka, N., Furuno, M., and Miyano, M. (1991) J. Biol. Chem., 266, 8426–8430.
Cho, H. J., Kim, S. J., and Kim, B. D. (2000) Mol. Cells, 10, 135–145.
Lord, J. M., Roberts, L. M., and Roberts, J. D. (1994) FASEB J., 8, 201–208.
Author information
Authors and Affiliations
Corresponding author
Additional information
__________
Translated from Biokhimiya, Vol. 70, No. 9, 2005, pp. 1218–1225.
Original Russian Text Copyright © 2005 by Gholizadeh, Baghban Kohnehrouz, Santha, Lodha, Kapoor.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM04-221, February 13, 2005.
Rights and permissions
About this article
Cite this article
Gholizadeh, A., Baghban Kohnehrouz, B., Santha, I.M. et al. Cloning and Expression of Small cDNA Fragment Encoding Strong Antiviral Peptide from Celosia cristata in Escherichia coli . Biochemistry (Moscow) 70, 1005–1010 (2005). https://doi.org/10.1007/s10541-005-0216-y
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/s10541-005-0216-y