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Judul Purification Of Recombinant Peritrophic Membrane Proteins Of The Old World Screwworm Fly, Chrysomya Bezziana. / Pearson, Roger; Muharsini, Sri[Research Institute for Veterinary Science]Wijffels, Gene; Vuocolo, Tony.
Pengarang Muharsini, Sri[Research Institute for Veterinary Science]Wijffels, Gene
Vuocolo, Tony.
EDISI Jurnal Ilmu Ternak dan Veteriner.
Penerbitan 2000
Deskripsi Fisik Vol.5 (3): p.185-191
Subjek Screwworm fly, Chrysomya bezziana, recombinant proteins, immobilized metal affinity chromatography
Abstrak To evaluate the feasibility of vaccinating sheep against the Old World Screwworm fly Chrysomya bezziana several recombinant peritrophin proteins were expressed in either a denatured form in Escherichia coli or a native-like form in Pichia pastoris cultures. Purification of the hexaHis tagged proteins was achieved by immobilized metal affinity chromatography. Proteins purified under reducing conditions were refolded using a glutathione shuffle procedure. Purification of a glutathione-Stransferase fusion protein was attempted using glutathione affinity chromatography in conjunction with anion exchange chromatography. The authenticity of the expressed proteins was verified by amino terminal amino acid sequencing. Carbohydrate analysis using biotinylated lectins revealed that Cb-peritrophin-48 expressed in Pichia pastoris was glycosylated with high mannose-type sugars. Four of the purified recombinant proteins were used to evaluate their protecti
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ARTVET2045 ARTVET2045 Dapat dipinjam Perpustakaan Balai Besar Pengujian Standar Instrumen Veteriner - Koleksi Agris Tersedia
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035 # # $a 0010-0721002231
082 # # $a ARTVET2045
084 # # $a ARTVET2045
245 # # $a Purification Of Recombinant Peritrophic Membrane Proteins Of The Old World Screwworm Fly, Chrysomya Bezziana. /$c Pearson, Roger; Muharsini, Sri[Research Institute for Veterinary Science]Wijffels, Gene; Vuocolo, Tony.
250 # # $a Jurnal Ilmu Ternak dan Veteriner.
260 # # ,$c 2000
300 # # $a Vol.5 (3): p.185-191
520 # # $a To evaluate the feasibility of vaccinating sheep against the Old World Screwworm fly Chrysomya bezziana several recombinant peritrophin proteins were expressed in either a denatured form in Escherichia coli or a native-like form in Pichia pastoris cultures. Purification of the hexaHis tagged proteins was achieved by immobilized metal affinity chromatography. Proteins purified under reducing conditions were refolded using a glutathione shuffle procedure. Purification of a glutathione-Stransferase fusion protein was attempted using glutathione affinity chromatography in conjunction with anion exchange chromatography. The authenticity of the expressed proteins was verified by amino terminal amino acid sequencing. Carbohydrate analysis using biotinylated lectins revealed that Cb-peritrophin-48 expressed in Pichia pastoris was glycosylated with high mannose-type sugars. Four of the purified recombinant proteins were used to evaluate their protective immunogenicity in sheep against Chrysomya bezziana strike.
650 # 4 $a Screwworm fly, Chrysomya bezziana, recombinant proteins, immobilized metal affinity chromatography
700 # $a Muharsini, Sri[Research Institute for Veterinary Science]Wijffels, Gene
700 # $a Vuocolo, Tony.
990 # # $a ARTVET2045
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