[Chinese little-fat-tail sheep prion protein gene belongs to PrPARH genotype].
The genomic DNA was extracted from lymphocytes of peripheral blood of Chinese little-fat-tail sheep. The prion protein gene(PrPc) was amplified by polymerase chain reaction. The PCR products were cloned into pUC19 plasmid and was sequenced. The results indicated that 771 bp amplified fragment contains the entire sheep PrPc coding sequence. There were 7 base pairs different with the reported sheep PrPc sequence, leading to three amino acids changes in the protein. It is show that its genotype is PrPARH, and these three polymorphisms may be linked to susceptibility or resistance to scrapie infection. Mature sheep prion protein gene was inserted into pET30a and expressed in E. coli, and 23 kD PrP25-231 protein was obtained. Recombinant PrP25-231 protein was purified by inclusion body washing, ion-exchange chromatography, reverse phase chromatography. Results of SDS-PAGE and Western blot showed that the purified PrP25-231 was obtained.
Li
Yun-Min
YM
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Tian
Bo
B
D000818:Animals; D015153:Blotting, Western; D015870:Gene Expression; D005838:Genotype; D010446:Peptide Fragments / Q000235:genetics / Q000302:isolation & purification; D011110:Polymorphism, Genetic; D011328:Prions / Q000235:genetics* / Q000302:isolation & purification; D012756:Sheep
D010446:Peptide Fragments; D011328:Prions