| 摘要 |
PURPOSE:To obtain the subject enzyme useful as a biochemical reagent making possible to fractionated determination of bile acid in affecting to the bile acid by culturing highly manifested colibacillus transformed with a gene- recombination method, extracting enzyme from said colibacillus and purifying. CONSTITUTION:Chromosome DNA of colibacillus is digested by restrictive enzyme BamHI and resultant DNA fragment is connected to BamHI-cutting position of colibacillus plasmid pBR322 to construct a fused plasmid, then resultant recombinant plasmid is introduced into colibacillus to produce a transformed strain, thus a highly manifesting transferring strain is prepared and resultant strain is subjected to screening, then cultured in a jar fermentor, thus bacterium is collected by centrifuge and the bacterium is crushed, then centrifuged, thus ammonium sulfate is added to resultant supernatant and precipitate at 60% saturation is separated, then resultant precipitate is dissolved in a buffer solution, thus purified by gel filtration, ion-exchange column chromatography and high-performance liquid chromatography to afford the aimed enzyme. |
