Chromatographic Technologic Research And Proteome Technical Application Of A New Tapy DNAase From Bimastos

This program have carried out systematic, overall chromato-graphic technologic research for this new tapy DNAase in earthworm organization; And through applying newest modern biological informatics and proteome technical system the partial informations and the structure of this new tapy DNAase are further appraised. 1. The chromatographic technologic research of this new tapy DNA -ase in earthworm organization.the enzyme vigour definition of DNAse I by Kunitz and Yasuda etc. can be as an active detection system of new DNA enzyme; By way of the specific property research of the physics and chemistry to this enzyme, the rudiment separation is prepared with high efficiency physics methods such as first through sugar dissolves and selectivity sour denaturation, the denaturation of thermal shock and ethanol precipitates etc. the most hybrid proteins are excised and a large number of target protein is fastly catched from the complex sample system; Then chromatographic technologic and separate character are studied by using the AKTA purifier of the company of Amersham Biosciences, Sepharose Fast Flow and Sephacryl series gel as well as Hitrap Selection Kit and XK series.result :The most suitable split packing for this enzyme is the weak anion exchange chromatography of Hitrap DEAE Sepharose Fast Flow, the hydrophobic interaction chromatography of Hitrap Phenyl Sepharose 6 Fast Flow ( low sub ) as well as sephacryl S-200. A new tapy DNAase sample of the size of 2. 4 Kda of the electrophore-sis level purity can be sparated. The the quick, efficient and stablereasonable refined purification technology route has been laieddown.2. The fast, accurate proteome technics application of this new tapyDNAase in earthworm organization.The PMF and the database of the peptides are gained by the tryptic digested in-gel, decolouring, the peptide drawing and Matrix mixing then through MALDI-TOF-MS. The target protein is appraised by MOWSE using mascot through SWISS-PROT database; The mass spectrum of MS/MS obtained by CID camepare with NCBI database.ResultrTwo sets of meaningful matching are gained through MOWSE:1. gi 543530, Alkaline trypsin-like serine proteinase (EC 3.4.21.-) F-I-0 - earthworm (Lumbricus rubellus) (fragment)2. gi | 3668171, RNA polymerase I second-largest subunit (Neurosp -ora crassa)retrieval shows one kind of new protein at last, the molecular weight is 24169. 34, ' s. retrieval shows what one has gained or acquired peptide section for the one kind of small molecule peptide inside the earthworm body and RNAase polymerase I from Neurospora crassa contain partially structure sequence function of Alkaline trypsin-like serine proteinase and RNAase polymerase I structure sequence function in this new protein.In a word, this program explore a fastly and efficiently stabilized shrewd purification technology successfully; At the same time the sample of purification be successfully appraised by using the bio-mass spectra technology of proteome.