杀虫晶体蛋白和低聚异麦芽糖的毛细管电泳分析-微生物学专业论文.docxVIP

AbstractThe Abstract The research results are summarized as following： Purification of crystal proteins(95％)by petroleum ether—extracted and aqueous two—phase extraction WaS described．Crude insecticidal crystal proteins(ICPs，133 kDa) was first dissolved in Na2C03 0H 1 0．5)，then precipitated in HCl(1 mol／L)and finally purified by DEAE ion-exchange chromatography．A capillary zone electrophoresis method and a capillary gel electrophoresis for determining ICPs in Bacillus thuringiensis were developed．With 50 mmol／L borax，3．5 mmol／L SDS，20％ACN and O．1％ p-mercaptoethanol at pH 9．5 as running buffer,ICPs catl be determined within 1 0 min at the applied voltage of 10 kV and the capillary temperature of 25C．The dimension of capillary Was 40 cm×75岫I．D without coating．The linear correlation coefficient between the concentration of ICPs and the peak area was equal to 0．9997 in the concentration range of 0．05～0．40 medrrd．The experiment conditions of capillary gel electrophoresis：neutrally-coated capillary(50 gm I．D，40 cm oftotal length and 30 cm of efficientlength)．0．1 mol／LTris．CHES(pH 8．6)，0．1％SDS，2．5％PEOas runningbuffer； the applied voltage of 300 V／cm；the capillary temperature at 25C and the detection wave—length Was 214 nln． A capillary eleetrophoresis method for the determination of main components in isomaltooligosaccharide formulation was described．The influences of variOIlS separartion conditions including buffer concentration，pH value，voltage and detection wave—length were investigated．By using an uncoated silica capillary(75 gm I．D，60 cm oftotal length， and 50 cm of efficient length)and 75 mmol／L borax 0H 1 0．5)as carrier electrolyte，the derivatlzed isomaltooligosaccharides with a—naphymethamine，including isomaltose， panose，isomaltotriose，wer@sufficiently separated at the qualification of an applied voltage of 15 kV and a UV detection wave．1ength of214 m．The results indicate that the relative standard deviations for migration times and peaks area were all less thaIl 0