Research Article
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Year 2018, Volume: 24 Issue: 2, 227 - 237, 01.06.2018
https://doi.org/10.15832/ankutbd.446442

Abstract

References

  • Abbas S (2007). Cloning and expression of cDNA encoding ribosome inactivating proteins. MSc Thesis, Dharwad University (Unpublished), India
  • Barbieri L & Stirpe F (1982). Ribosome-inactivating proteins from plants: Properties and possible uses. Cancer Survey 1: 489-520
  • Barbieri L, Battelli M G & Stirpe F (1993). Ribosome inactivating proteins from plants. Biochimica et Biophysica Acta 1154: 237-282
  • Barbieri L, Polito L, Bolognesi A, Ciani M, Pelosi E, Farini V, Jha A K, Sharma N, Vivanco J M, Chambery A, Parente A & Stirpe F (2006). Ribosomeinactivating proteins in edible plants and purification and characterization of a new ribosome-inactivating protein from Cucurbita moschata. Biochimica et Biophysica Acta 1760: 783-792
  • Bolognesi A, Polito L, Olivieri F, Valbonesi P, Barbieri L, Giulia Battelli M, Vittoria Carusi M, Benvenuto E, Del Vecchio Blanco F, Di Maro A, Parente A, Di Loreto M & Stirpe F (1997). New ribosome inactivating proteins with polynucleotide: adenosine glycosidase and antiviral activities from Basella rubra L. and Bougainvillea spectabilis Willd. Planta 203: 422-429
  • Bolognesi A, Polito L, Tazzari P L, Lemoli R M, Lubelli C, Fogli M, Boon L, de Boer M & Stirpe F (2000). In vitro anti-tumour activity of anti-CD80 and antiCD86 immunotoxins containing type 1 ribosomeinactivating proteins. British Journal of Haematology 110: 351-361
  • Bradford M M (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254
  • Choudhary N L, Yadav O P & Lodha M L (2007). Ribonuclease, deoxyribonuclease, and antiviral activity of Escherichia coli-expressed Bougainvillea xbuttiana antiviral protein. Biochemistry (Moscow) 73: 273-277
  • Choudhary N, Kapoor H C & Lodha M L (2008). Cloning and expression of antiviral/ribosome-inactivating protein from Bougainvillea xbuttiana. Journal of Biosciences 33: 91-101
  • Christopher G W, Cieslak T J Z, Pavlin J A & Eitzen E M Jr (1997). Biological warfare. A historical perspective. Jama 278: 412-417
  • Donayre Torres A J, Esquivel Soto E, Gutierrez Xicotencatl M D, Esquivel Guadarrama F R & Gomez Lim M A (2009). Production and purification of immunologically active core protein p24 from HIV-1 fused to ricin toxin B subunit in E. coli. Journal of Virology 6: 1-11
  • Endo Y & Tsurugi K (1988). The RNA N-glycosidase of ricin A chain: The characteristics of enzymatic activity of ricin A chain with ribosomes and rRNA. The Journal of Biological Chemistry 263: 8735-8739
  • Endo Y, Mitsui K, Motizuki M & Tsurugi K (1987). The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. The site and the characteristics of the modification in 28 S ribosomal RNA caused by the toxins. Journal of Biological Chemistry 262(12): 5908-5912
  • Foissac L, Gentit P, Svanetia-Dumas L, Dulucq M J & Candresse T (2001). Polyvalent detection of fruit tree tricho, capillo, and fovea viruses by nested RT-PCR using degenerated and inosine containing primers (PDO RT-PCR). Acta Horticulturae 550: 37-43
  • Gessner S L & Irvin J D (1980). Inhibition of elongation factor2-dependent translocation by the pokeweed antiviral protein and ricin. Journal of Biological Chemistry 255: 3251-3253
  • Girbés T, Ferreras J M, Arias F J & Stirpe F (2004). Description, distribution, activity and phylogenetic relationship of ribosome inactivating proteins in plants, fungi and bacteria. Mini-Reviews in Medicinal Chemistry 4: 461-476
  • Grasso S & Shepherd R J (1978). Isolation and partial characterization of virus inhibitors from plant species taxonomically related to Phytolacca. Journal of Physiology Biochemistry 68: 199-205
  • Hakuba N, Miyano M, Kataoka J & Noma M (1991). Escherichia coli ribosome is inactivated by Mirabilis antiviral protein which cleaves the N-glikosidic bond at A2660 of 23S ribosomal RNA. Journal of Molecular Biology 221: 737-743
  • Hartley M R, Legname G, Osborn R, Chen Z & Lord J M (1991). Single chain ribosome inactivating proteins from plants depurinate Escherichia coli 23S ribosomal RNA. FEBS Letters 290(1-2): 65-68
  • Hartley M R, Chaddock J A & Bonness M S (1996). The structure and function of ribosome-inactivating proteins. Trends in Plant Science 1: 254-260 Irvin J D (1983). Pokeweed antiviral protein. Pharmacology & Therapeutics 21: 371-387
  • Irvin J D (1995). Antiviral proteins from Phytolacca. In: M Chessin, A Zipf & D DeBorde (Eds), Antiviral Proteins from Higher Plants. CRC Press, Boca Raton, FL pp. 65-69
  • Jackman M R, Ellis J A, Gray S R, Shurety W & Luzio J P (1999). Cell polarization is required for ricin sensitivity in a Caco-2 cell line selected for ricin resistance. Biochemistry Journal 341: 323-327
  • Kobayashi K D, McConnell J & Griffis J (2007). Bougainvillea. Ornamentals and Flowers 38: 1-12
  • Kwon S Y, An C S, Liu J R, Kwak S S, Lee H S, Lim J K & Paek K H (2000). Molecular cloning of a cDNA encoding ribosome-inactivating protein from Amaranthus viridis and its expression in E. coli. Molecules and Cells 10: 8-12
  • Laemmli U K (1970). Cleavage of structural proteins during the assembly of the head of bacteriophasge T4. Nature 227: 680-685
  • Liu L, Kloepper J W & Tuzun S (1995). Introduction of systemic resistance in cucumber against Fusarium wilt by plant growth-promoting rhizobacteria. Phytopathology 85: 695-698
  • Lodge J K, Kaniewski W K & Tumer N E (1993). Broad spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein. Proceedings of the National Academy of Sciences of the United Stated of America 90: 7089-7093
  • Lodha M L, Agarwal S, Biswas K, Vasudev S & Dubey S C (2010). Antimicrobial activity of native and recombinant antiviral proteins from Bougainvillea xbuttiana leaves against plant pathogenic fungi and viruses. Indian Journal of Agricultural Biochemistry 23: 83-90
  • Logemann J, Jach G, Tommerup H, Mundy J & Schell J (1992). Expression of a barley ribosome-inactivating protein leads to increased fungal protection in transgenic tobacco plants. Biotechnology 10: 305-308
  • Madin K, Saeasaki T, Ogasawara T & Endo Y (2000). A highly efficient and robust cell-free protein synthesis system prepared from wheat embryos: Plants apparently contain a suicide system directed at ribosomes. Proceedings of the National Academy of Science of the United Stated of America 97: 559-564
  • Nielsen K, Payne G A & Boston R S (2001). Maize ribosome inactivating protein inhibits normal development of Aspergillus nidulans and Aspergillus flavus. Molecular Plant-Microbe Interactions Journal 14: 164-172
  • Olsnes S & Pihl A (1980). Toxic lectins and related proteins. In: P H Cohen & S van Heyningen (Eds), The Molecular Actions of Toxins and Viruses, Elsevier/ North Holland, Amsterdam, pp. 51-106
  • Özer M, Sipahioğlu H M, Usta M & Fidan H (2012). Cloning and sequencing of coat protein gene of Zucchini yellow mosaic virus isolated from squash and muskmelon in Turkey. Turkish Journal of Biology 36: 423-429
  • Qi L, Nett T M, Allen M C, Sha X, Harrison G S, Frederick B A, Crawford E D & Glode L M (2004). Binding and cytotoxicity of conjugated and recombinant fusion proteins targeted to the gonadotropin-releasing hormone receptor. Cancer Research 64: 2090-2095
  • Roberts W K & Selitrennikoff C P (1986). Isolation and characterization of two antifungal proteins from barley. Biochimica et Biophysica Acta 880: 161-170
  • Sadasivam S, Rajameshewari S & Jeyarajan R (1991). Inhibition of certain plant viruses by plant extracts. Journal of Ecobiology 3: 53-57
  • Sankian M, Yousefi M, Pazouki N & Varasteh A (2007). One-step purification of histidine-tagged profilin with high purity and yield by using metal precipitation. Biotechnology and Applied Biochemistry 47: 185-189
  • SAS (2014). SAS/STAT Software: Hangen and Enhanced. SAS, Inst. Inc., Cary, N.C. USA Scadden D T, Schenkein D P, Bernstein Z, Luskey B, Doweiko J, Tulpule A & Levine A M (1998). Immunotoxin combined with chemotherapy for patients with AIDS-related nonHodgkin’s lymphoma. Cancer 83: 2580-2587
  • Schlumbaum A, Mauch F, Vogeli U & Boller T (1986). Plant chitinases are potent inhibitors of fungal growth. Nature 324: 365-367
  • Schmitt A C, Ravazzolo A P & Von Poser G L (2001). Investigation of some Hypericum species native to Southern of Brazil for antiviral activity. The Journal of Ethnopharmacology 77: 239-245
  • Schnell R, Katouzi A A, Linnartz C, Schoen G, Drillich S, Hansmann M L, Schiefer D, Barth S, ZangemeisterWittke U, Stahel R A, Diehl V & Engert A (1996). Potent anti-tumor effects of an anti-CD24 ricin A-chain immunotoxin in vitro and in a disseminated human Burkitt’s lymphoma model in SCID mice. International Journal of Cancer 66: 526-531
  • Stevens W A, Spurdon C, Onyon L J & Stirpe F (1981). Effect of inhibitors of protein synthesis from plants on tobacco mosaic virus infection. Experientia 37: 257-259
  • Stirpe F & Barbieri L (1986). Ribosome-inactivating proteins up to date. FEBS Letters 195: 1-8
  • Stirpe F, Gasperi-Campani A, Barnieri L, Falasca A, Abbondanza A & Stevens W A (1983). Ribosomeinactivating proteins from the seeds of Saponara officinalis L. (soapwort), of Agrostemma githago L. (corn cockle) and of Asparagus officinalis L. (asparagus), and from the latex of Hura crepitans L. (sandbox tree). Biochemical Journal 216: 617-625
  • Stirpe F, Barbieri L, Battelli M G, Soria M & Lappi D A (1992). Ribosome-inactivating proteins from plants: present status and future prospects. Biotechnology 10: 405-412
  • Wang P, Zoubenko O & Tumer N E (1998a). Reduced toxicity and broad spectrum resistance to viral and fungal infection in transgenic plants expressing pokeweed antiviral protein II. Plant Molecular Biology 38: 957-964
  • Wang W X, Dong J Y, Zhou S Y, Li W L & Zhao Y (1998b). Modification of ricin and its hepatotoxicity and activity against hepatocellular cancer in mice. World Journal Gastroenterology 4: 307-310
  • Weiner S L (1996). Strategies for the prevention of a successful biological warfare aerosol attack. Military Medicine 161: 251-256

Antiviral and Antifungal Activity of Biologically Active Recombinant Bouganin Protein from Bougainvillea spectabilis Willd

Year 2018, Volume: 24 Issue: 2, 227 - 237, 01.06.2018
https://doi.org/10.15832/ankutbd.446442

Abstract

Bouganin antiviral protein (BAP) gene, one of the ribosome inactivating proteins, isolated from Bougainvillea spectabilis Willd. was cloned, expressed and the antiviral and antifungal activities were investigated. The full-length bouganin antiviral protein gene was amplified by reverse transcription-PCR using mRNA as template extracted from mature leaves. The coding region of bouganin gene was cloned into prokaryotic expression vector pETDuet-1 after amplification with end to end gene specific primers. The recombinant plasmid was transformed into Escherichia coli cells BL21(DE3)pLysS and the expression of BAP gene was induced by isopropyl β-D thiogalactopyranoside (IPTG). Bouganin antiviral protein having a molecular mass of 28 kDa has been isolated from transformed bacterial colonies. Antiviral activity of bouganin was assayed against Zucchini yellow mosaic virus (ZYMV) by a mechanical inoculation test. The antifungal activity of purified recombinant protein was tested against pathogenic and non-pathogenic Rhizoctonia solani, Trichoderma harzianum, and Fusarium oxysporum fungi using disc diffusion method. The increased amount of antiviral protein reduced the disease severity caused by ZYMV. The bouganin antiviral protein was inhibited the growth of R. solani by 30.7% and of T. harzianum by 20% after 72 h compared to control. No growth inhibition was observed for F. oxyporum. All plants including controls treated with in vitro expressed BAP protein exhibited severe growth reduction compared with negative control (not treated) plants. 


References

  • Abbas S (2007). Cloning and expression of cDNA encoding ribosome inactivating proteins. MSc Thesis, Dharwad University (Unpublished), India
  • Barbieri L & Stirpe F (1982). Ribosome-inactivating proteins from plants: Properties and possible uses. Cancer Survey 1: 489-520
  • Barbieri L, Battelli M G & Stirpe F (1993). Ribosome inactivating proteins from plants. Biochimica et Biophysica Acta 1154: 237-282
  • Barbieri L, Polito L, Bolognesi A, Ciani M, Pelosi E, Farini V, Jha A K, Sharma N, Vivanco J M, Chambery A, Parente A & Stirpe F (2006). Ribosomeinactivating proteins in edible plants and purification and characterization of a new ribosome-inactivating protein from Cucurbita moschata. Biochimica et Biophysica Acta 1760: 783-792
  • Bolognesi A, Polito L, Olivieri F, Valbonesi P, Barbieri L, Giulia Battelli M, Vittoria Carusi M, Benvenuto E, Del Vecchio Blanco F, Di Maro A, Parente A, Di Loreto M & Stirpe F (1997). New ribosome inactivating proteins with polynucleotide: adenosine glycosidase and antiviral activities from Basella rubra L. and Bougainvillea spectabilis Willd. Planta 203: 422-429
  • Bolognesi A, Polito L, Tazzari P L, Lemoli R M, Lubelli C, Fogli M, Boon L, de Boer M & Stirpe F (2000). In vitro anti-tumour activity of anti-CD80 and antiCD86 immunotoxins containing type 1 ribosomeinactivating proteins. British Journal of Haematology 110: 351-361
  • Bradford M M (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254
  • Choudhary N L, Yadav O P & Lodha M L (2007). Ribonuclease, deoxyribonuclease, and antiviral activity of Escherichia coli-expressed Bougainvillea xbuttiana antiviral protein. Biochemistry (Moscow) 73: 273-277
  • Choudhary N, Kapoor H C & Lodha M L (2008). Cloning and expression of antiviral/ribosome-inactivating protein from Bougainvillea xbuttiana. Journal of Biosciences 33: 91-101
  • Christopher G W, Cieslak T J Z, Pavlin J A & Eitzen E M Jr (1997). Biological warfare. A historical perspective. Jama 278: 412-417
  • Donayre Torres A J, Esquivel Soto E, Gutierrez Xicotencatl M D, Esquivel Guadarrama F R & Gomez Lim M A (2009). Production and purification of immunologically active core protein p24 from HIV-1 fused to ricin toxin B subunit in E. coli. Journal of Virology 6: 1-11
  • Endo Y & Tsurugi K (1988). The RNA N-glycosidase of ricin A chain: The characteristics of enzymatic activity of ricin A chain with ribosomes and rRNA. The Journal of Biological Chemistry 263: 8735-8739
  • Endo Y, Mitsui K, Motizuki M & Tsurugi K (1987). The mechanism of action of ricin and related toxic lectins on eukaryotic ribosomes. The site and the characteristics of the modification in 28 S ribosomal RNA caused by the toxins. Journal of Biological Chemistry 262(12): 5908-5912
  • Foissac L, Gentit P, Svanetia-Dumas L, Dulucq M J & Candresse T (2001). Polyvalent detection of fruit tree tricho, capillo, and fovea viruses by nested RT-PCR using degenerated and inosine containing primers (PDO RT-PCR). Acta Horticulturae 550: 37-43
  • Gessner S L & Irvin J D (1980). Inhibition of elongation factor2-dependent translocation by the pokeweed antiviral protein and ricin. Journal of Biological Chemistry 255: 3251-3253
  • Girbés T, Ferreras J M, Arias F J & Stirpe F (2004). Description, distribution, activity and phylogenetic relationship of ribosome inactivating proteins in plants, fungi and bacteria. Mini-Reviews in Medicinal Chemistry 4: 461-476
  • Grasso S & Shepherd R J (1978). Isolation and partial characterization of virus inhibitors from plant species taxonomically related to Phytolacca. Journal of Physiology Biochemistry 68: 199-205
  • Hakuba N, Miyano M, Kataoka J & Noma M (1991). Escherichia coli ribosome is inactivated by Mirabilis antiviral protein which cleaves the N-glikosidic bond at A2660 of 23S ribosomal RNA. Journal of Molecular Biology 221: 737-743
  • Hartley M R, Legname G, Osborn R, Chen Z & Lord J M (1991). Single chain ribosome inactivating proteins from plants depurinate Escherichia coli 23S ribosomal RNA. FEBS Letters 290(1-2): 65-68
  • Hartley M R, Chaddock J A & Bonness M S (1996). The structure and function of ribosome-inactivating proteins. Trends in Plant Science 1: 254-260 Irvin J D (1983). Pokeweed antiviral protein. Pharmacology & Therapeutics 21: 371-387
  • Irvin J D (1995). Antiviral proteins from Phytolacca. In: M Chessin, A Zipf & D DeBorde (Eds), Antiviral Proteins from Higher Plants. CRC Press, Boca Raton, FL pp. 65-69
  • Jackman M R, Ellis J A, Gray S R, Shurety W & Luzio J P (1999). Cell polarization is required for ricin sensitivity in a Caco-2 cell line selected for ricin resistance. Biochemistry Journal 341: 323-327
  • Kobayashi K D, McConnell J & Griffis J (2007). Bougainvillea. Ornamentals and Flowers 38: 1-12
  • Kwon S Y, An C S, Liu J R, Kwak S S, Lee H S, Lim J K & Paek K H (2000). Molecular cloning of a cDNA encoding ribosome-inactivating protein from Amaranthus viridis and its expression in E. coli. Molecules and Cells 10: 8-12
  • Laemmli U K (1970). Cleavage of structural proteins during the assembly of the head of bacteriophasge T4. Nature 227: 680-685
  • Liu L, Kloepper J W & Tuzun S (1995). Introduction of systemic resistance in cucumber against Fusarium wilt by plant growth-promoting rhizobacteria. Phytopathology 85: 695-698
  • Lodge J K, Kaniewski W K & Tumer N E (1993). Broad spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein. Proceedings of the National Academy of Sciences of the United Stated of America 90: 7089-7093
  • Lodha M L, Agarwal S, Biswas K, Vasudev S & Dubey S C (2010). Antimicrobial activity of native and recombinant antiviral proteins from Bougainvillea xbuttiana leaves against plant pathogenic fungi and viruses. Indian Journal of Agricultural Biochemistry 23: 83-90
  • Logemann J, Jach G, Tommerup H, Mundy J & Schell J (1992). Expression of a barley ribosome-inactivating protein leads to increased fungal protection in transgenic tobacco plants. Biotechnology 10: 305-308
  • Madin K, Saeasaki T, Ogasawara T & Endo Y (2000). A highly efficient and robust cell-free protein synthesis system prepared from wheat embryos: Plants apparently contain a suicide system directed at ribosomes. Proceedings of the National Academy of Science of the United Stated of America 97: 559-564
  • Nielsen K, Payne G A & Boston R S (2001). Maize ribosome inactivating protein inhibits normal development of Aspergillus nidulans and Aspergillus flavus. Molecular Plant-Microbe Interactions Journal 14: 164-172
  • Olsnes S & Pihl A (1980). Toxic lectins and related proteins. In: P H Cohen & S van Heyningen (Eds), The Molecular Actions of Toxins and Viruses, Elsevier/ North Holland, Amsterdam, pp. 51-106
  • Özer M, Sipahioğlu H M, Usta M & Fidan H (2012). Cloning and sequencing of coat protein gene of Zucchini yellow mosaic virus isolated from squash and muskmelon in Turkey. Turkish Journal of Biology 36: 423-429
  • Qi L, Nett T M, Allen M C, Sha X, Harrison G S, Frederick B A, Crawford E D & Glode L M (2004). Binding and cytotoxicity of conjugated and recombinant fusion proteins targeted to the gonadotropin-releasing hormone receptor. Cancer Research 64: 2090-2095
  • Roberts W K & Selitrennikoff C P (1986). Isolation and characterization of two antifungal proteins from barley. Biochimica et Biophysica Acta 880: 161-170
  • Sadasivam S, Rajameshewari S & Jeyarajan R (1991). Inhibition of certain plant viruses by plant extracts. Journal of Ecobiology 3: 53-57
  • Sankian M, Yousefi M, Pazouki N & Varasteh A (2007). One-step purification of histidine-tagged profilin with high purity and yield by using metal precipitation. Biotechnology and Applied Biochemistry 47: 185-189
  • SAS (2014). SAS/STAT Software: Hangen and Enhanced. SAS, Inst. Inc., Cary, N.C. USA Scadden D T, Schenkein D P, Bernstein Z, Luskey B, Doweiko J, Tulpule A & Levine A M (1998). Immunotoxin combined with chemotherapy for patients with AIDS-related nonHodgkin’s lymphoma. Cancer 83: 2580-2587
  • Schlumbaum A, Mauch F, Vogeli U & Boller T (1986). Plant chitinases are potent inhibitors of fungal growth. Nature 324: 365-367
  • Schmitt A C, Ravazzolo A P & Von Poser G L (2001). Investigation of some Hypericum species native to Southern of Brazil for antiviral activity. The Journal of Ethnopharmacology 77: 239-245
  • Schnell R, Katouzi A A, Linnartz C, Schoen G, Drillich S, Hansmann M L, Schiefer D, Barth S, ZangemeisterWittke U, Stahel R A, Diehl V & Engert A (1996). Potent anti-tumor effects of an anti-CD24 ricin A-chain immunotoxin in vitro and in a disseminated human Burkitt’s lymphoma model in SCID mice. International Journal of Cancer 66: 526-531
  • Stevens W A, Spurdon C, Onyon L J & Stirpe F (1981). Effect of inhibitors of protein synthesis from plants on tobacco mosaic virus infection. Experientia 37: 257-259
  • Stirpe F & Barbieri L (1986). Ribosome-inactivating proteins up to date. FEBS Letters 195: 1-8
  • Stirpe F, Gasperi-Campani A, Barnieri L, Falasca A, Abbondanza A & Stevens W A (1983). Ribosomeinactivating proteins from the seeds of Saponara officinalis L. (soapwort), of Agrostemma githago L. (corn cockle) and of Asparagus officinalis L. (asparagus), and from the latex of Hura crepitans L. (sandbox tree). Biochemical Journal 216: 617-625
  • Stirpe F, Barbieri L, Battelli M G, Soria M & Lappi D A (1992). Ribosome-inactivating proteins from plants: present status and future prospects. Biotechnology 10: 405-412
  • Wang P, Zoubenko O & Tumer N E (1998a). Reduced toxicity and broad spectrum resistance to viral and fungal infection in transgenic plants expressing pokeweed antiviral protein II. Plant Molecular Biology 38: 957-964
  • Wang W X, Dong J Y, Zhou S Y, Li W L & Zhao Y (1998b). Modification of ricin and its hepatotoxicity and activity against hepatocellular cancer in mice. World Journal Gastroenterology 4: 307-310
  • Weiner S L (1996). Strategies for the prevention of a successful biological warfare aerosol attack. Military Medicine 161: 251-256
There are 48 citations in total.

Details

Primary Language English
Subjects Engineering
Journal Section Makaleler
Authors

Abdullah Güller This is me

Hikmet Murat Sipahioğlu

Mustafa Usta This is me

Emre Demirer Durak This is me

Publication Date June 1, 2018
Submission Date November 25, 2016
Acceptance Date March 30, 2017
Published in Issue Year 2018 Volume: 24 Issue: 2

Cite

APA Güller, A., Sipahioğlu, H. M., Usta, M., Durak, E. D. (2018). Antiviral and Antifungal Activity of Biologically Active Recombinant Bouganin Protein from Bougainvillea spectabilis Willd. Journal of Agricultural Sciences, 24(2), 227-237. https://doi.org/10.15832/ankutbd.446442

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