摘要
寻找大肠杆菌E.coli脂多糖的最佳提取方法。分别用改良热酚法、超声波处理法、煮沸法从大肠杆菌细胞壁中提取脂多糖,经纯化后分别在721分光光度仪上测其吸光值;再用3种方法提取的脂多糖分别免疫草鱼,计数法测受试草鱼血液中白细胞的数量,愈创木酚法测受试鱼不同组织中过氧化氢酶的活力(用OD值表示)。改良热酚法、超声波处理法、煮沸法提取的脂多糖在稀释15倍时,其吸光值分别是1.973、1.003和1.153;改良热酚法、超声波处理法、煮沸法提取的脂多糖在稀释15倍时,免疫草鱼测得受试鱼的白细胞数量分别为159个/μL、160个/μL和135个/μL,测得受试鱼血液中的过氧化氢酶活力分别是0.09、0.083、0.078,测得脾脏中的过氧化氢酶活力分别是0.083、0.078、0.06。改良热酚法提取的大肠杆菌脂多糖浓度高、免疫活性强,是相对比较好的一种提取方法。
To study and optimize the extracting technology of the lipopolysaccharide of E.coli.The improved hot phenol method,ultrasonic treatment,boiling were used to extract the lipopolysaccharide from the E.coli cell wall,and the products of three methods were purified and detected the absorption value with 721 spectrophotometer,and then were differently injected the grass carp,tested the number of white corpuscles in grass carp blood and catalase activity in different the tissue of grass carp with guaiacol method （data shown with OD value）.The lipopolysaccharide extracted with the improved hot phenol,ultrasonic and boiling method at diluted 15-folds was separately detected OD value （1.973,1.003 and 1.153） and was injected the grass carp.The measured number of white corpuscles was 159 cells/μ L,160 cells/μL and 135 cells/μ L,respectively,the catalase activity in blood was 0.09,0.083 and 0.078,in spleen 0.083,0.078,and 0.06,respectively.The improved hot phenol method was suitable to extract the high concentrate and immune activity lipopolysaccharide.
出处
《中国农学通报》
CSCD
北大核心
2010年第21期12-15,共4页
Chinese Agricultural Science Bulletin
基金
河南省科技厅基础与前沿(102300410207)
河南省教育厅(2007180038)
