Sheep pox viral envelope protein L1RΔ synthesis in plants
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Keywords:
sheep pox virus, transgenic plants,Abstract
Sheep pox viral gene SPPV-NISKHI-56 encodes ortholog of immunodominant protein L1R of vaccinia virus. Gene sequence that encodes hydrophilic part of the protein (L1RΔ) was artificially synthesized and cloned in pET-19b plasmid for expression in bacteria. Recombinant DNA – construct based on pCAMBIA2300 vector was designed for transformation of the tobacco plants (Nicotiana tabacum L. cv. Samsun-NN). This construct contained 35S promoter of cauliflower mosaic virus, 5´UTR of gRNA AMV, His-Tag, gene for L1RΔ protein and NOS terminator. Transformed regenerated plants that produced root on the medium with kanamycin were analyzed for the presence of necessary mRNA by RT-PCR. Recombinant protein was detected in several transgenic plants by specific antibodies for L1RΔ.References
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5 Hooper J.W., Custer D.M., Schmaljohn C.S., Schmaljohn A.L. DNA vaccination with vaccinia virus L1R and A33R genes protects mice against a lethal poxvirus challenge // Virology. – 2000. – V. 266. – P. 329–339.
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9 Dilworth E., Frey J.E. A rapid method for high throughput DNA extraction from plant material for PCR amplification // Plant Molecular Biology Reporter. – 2000. – V. 18(1). – P. 61-64.
10 Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding // Analytical Biochemistry. – 1976. – V. 72. – P. 248-254.
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Beisenov, D., Stanbekova, G., Nadirova, L., & Iskakov, B. (2015). Sheep pox viral envelope protein L1RΔ synthesis in plants. Experimental Biology, 60(2), 187–190. Retrieved from https://bb.kaznu.kz/index.php/biology/article/view/157
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BIOTECHNOLOGY
