Establishment Of Two-dimensional Electrophoresis System And Proteome Analysis Of Sika Deer Antler

Sika deer are at the top of animal medicines, they play an irreplaceable role intraditional Chinese medicine materials because of their magical medicinal value andnourishing function. The compositions were unknown which restricted the development ofantlers industry. Meanwhile, the imperfection of antlers quality standard system led to thedecrease of competitive power in the world. So, how to analyze compositions and establishquality standard of deer antlers by molecular biology methods became an imperativeproblem for the development of sika deer industry. In this thesis, we sieved an optimalmethod for total proteins extraction, established2-DE system and analyzed proteome of sikadeer antlers, which can supply theory evidence for antlers production development.Firstly, we build a logical2-DE process of sika deer antler. The main steps were toestablish optimal total proteins extraction method and set up the correct equipmentparameter when focused, with the purpose of get a clear gel map. Results showed thatmanual lysate was the suitable method for total protein extraction which can get highconcentration sample and better protein map with more plain spots and less stripes.Secondly, PDQuest software was used to detect spots when proteome analysis. The cutinterest spots were identified by MALDI-TOF/TOF MS technology, and results weresearched in NCBI database. Finally, we judged if a spot was identified successfully or notby protein score. The next task was to define functions to find proteins related to growth andhealthcare. Results showed that460spots were detected and177individual proteins wereidentified, including31structural proteins,65enzymes,5inhibitor and activator factors,30signaling and channel proteins,3immune proteins,13molecular chaperones,14proteinsrelated to replication, transcription and translation,10blood proteins,6tumorigenesisproteins.At last24functional proteins were sieved.14of these proteins were related to growth,including centractin, vimentin, ATPase, alpha-enolase, pyruvate kinase, mitochondrial citratesynthase, malate dehydrogenase, fumarate hydratase, alcohol dehydrogenase, myoglobin,fibrinogen, EEF protein, pre-mRNA-processing factor, tRNA synt. In addition, heat shockproteins were related to wound healing and tenascin on nerve regeneration. Calreticulin,collagen, caldesmon, SPARC, reticulocalbin, FK506binding protein were engaged inossification. Superoxide dismutase (Mn) and peroxiredoxin were supposed to retard senility.