Effects Of Bovine Brain Tissue Intakes On The Ability Of Learning And Memory And The Structural And Functional Study Of Prion Protein In Mice

Since the 1960s, It is widely acknowledged that brain of bovine contains a rich amount of protein, lipid, vitarmin and trace elements as potassium; Improvement of and neurological impaired the prognosis memory in animal by bovine brain tissue intakes.However, Recent research indicates that ganglioside, trans fatty acide, myelin base protein, peripheral nerve myelin, hot HSP104 rush proteins, PrPC and correlated factor expression can have serious consequence for the health of human beings. In order to investigate whether the bovine brain tissue ingredients play bad effects on mammalian learning and memory ability and related mechanism, BSE free bovine brain tissue was prepared into special feeds and digested in a simulated human gastrointestinal tract environment to obtain active extracts. One hundred and twenty mice in four groups were fed with special feeds and or intraperitoneal injected with active brain extracts for 5 months, including experimental controls.GroupⅠmice were fed feed contains bovine brain;GroupⅡmice were fed feed contains bovine brain and intraperitoneal injection of bovine brain extract. GroupⅢmice were fed with a standard diet and pump digestive fluids.GroupⅣmice were fed with standard diet.By the end of this test.this paper has completed the following research work to effect of bovine brain tissue intakes on ability of learning and memory in mice and cloning and sequence analysis of prion protein gene in mice, Glycosylation and misfolding of PrP, the results are showed as follows:1. Effect of bovine brain tissue intakes on ability of learning and memory in mice.The ability of learning and memory were tested on all mice with water maze in the end of 30d,90d and 150d. The results showed that expression of the derived neurophic factor was reduced as well as the ability of learning and memory among the feeding or injecting groups. Thus, it revealed the bad effects of long-term bovine tissue intakes played on the mice.2. Cloning of PRNP gene and Prokaryotic expression and purification of Prion protein. Extraction of Genomic DNA in brain and expression vector PrP/pET-32 was constructed, the recombinant pMD19 T-adiponectin plasmid was digestes into two fragments by EcoR I and BamHⅠ, the DNA of PRNP was cloned from PRNP gene by polymerase chain reaction(PCR) and was inserted into pET-32 plasmid by using T4 DNA ligase. The sequenced DNA fragment was 840bp long, and it possessed a substitution at amino acis residue 297, a substitution at amino acis residue 300, a substitution at amino acis residue 300. a new protein band was found in SDS-PAGE with molecular mass of about 37Ku which is consisted of a 32Ku protein.3. Effect of bovine brain tissue intakes on PrPc structure and functions in PrPc. PrPsc in the brains were detected by immunohistochemistry. The test result revealed that all groups were not emerged claret colour particles in brain cinerea. The levels of PrPC of glycosylation were detected by immunoblotting. The ecpressed products were digested with PNGase F. prion protein of glycosylation sites mutated were down. Resulting in their molecular weights reduced, while the molecular weights were 27Ku. Mono-or non-N-glycosylation had been obtained successfully in the study. Gly-protein in examination of brain had been changed among non-glycosylated and one-N-glycosylated and two N-glycosylation sites why has the change taken place, the glycation might occur in vivo or vitro, Which could be a useful means for studying prions.