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Cloning, Sequencing, and Expression of cDNA Encoding Bovine Prion Protein  

Kang, Sang-Gyun (Department of Infectious Disease, Theriogenology ,College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
Kang, Sung-Keun (Department of Theriogenology and Biotechnology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
Lee, Deog-Yong (Department of Infectious Disease, Theriogenology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
Park, Yong-Ho (Department of Infectious Disease, Microbiology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
Hwang, Woo-Suk (Department of Infectious Disease and Biotechnology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
Yoo, Han-Sang (Department of Infectious Disease, Theriogenology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
Publication Information
Journal of Microbiology and Biotechnology / v.14, no.2, 2004 , pp. 417-421 More about this Journal
Abstract
A normal prion protein (PrPc) is converted to a protease resistant isoform (PrPsc) by an apparent self-propagating activity in bovine spongiform encephalopathies (BSE), which is a neurodegenerative disease. The cDNA encoding bovine PrP open reading frame (ORP) in Korean cattle was cloned by polymerase chain reaction (PCR). The cloned cDNA had a length of 795 base pairs which coded for a protein of 264 amino acid residues with a calculated molecular mass of 28.6 kDa. Identities of 90, 90, 79 and 78% on nucleotide and 94, 94, 84, and 84% on amino acid sequence were shown to PrP genes from sheep, goat, human, and mouse, respectively. The cloned DNA was ligated into the pQE30 expression vector and transformed into E. coli M15. The PrP was expressed by induction with isopropyl-$\beta$-D-thiogalactoside (IPTG) and purified on the Ni-NTA affinity column. High specific activities of the recombinant PrP were observed in the fraction of pH 5.8 eluate and showed a molecular mass of-29 kDa on SDS-PAGE and Western blot analysis.
Keywords
Prien protein; cDNA; BSE; Korean cattle;
Citations & Related Records
Times Cited By KSCI : 3  (Citation Analysis)
Times Cited By Web Of Science : 5  (Related Records In Web of Science)
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