丙型副伤寒沙门氏菌脂多糖提取纯化及活性分析  

Extraction,Purification and Activity Analysis of Lipopolysaccharide of Salmonella paratyphi C

作　　者：杨成兰[1] 罗薇[1] 刘内生[1] 杨阳[1] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041

出　　处：《中国畜牧兽医》2014年第4期75-78,共4页China Animal Husbandry & Veterinary Medicine

基　　金：四川省应用基础研究计划项目(2013JY0045)

摘　　要：为了提取、纯化天鹅源丙型副伤寒沙门氏菌脂多糖（LPS）并检测其活性，试验通过热酚水法提取丙型副伤寒沙门氏菌LPS，采用DNaseI、RNaseA和蛋白酶K及醇沉法纯化LPS，测定LPS提取物中多糖、蛋白及核酸的含量，鲎试剂检测凝集活性，显色基质法测定其活性。结果显示，纯化的丙型副伤寒沙门氏菌LPS平均产率为1．48％，多糖含量为3．84％，蛋白含量为1．49％，核酸含量为5．45％，且核酸片段低于100bp，SDS-PAGE电泳和银染结果显示条带主要集中在10～15ku范围内，与2EU／mL鲎试剂的最小凝集浓度是10．99ng／mL，显色基质法测定其活性为9．82×10^5EU／mg。该试验提取、纯化的丙型副伤寒沙门氏菌LPS纯度较高，生物活性良好。This study was to obtain lipopolysaccharide （LPS） from Salmonella paratyphi C and analyze its activity. In this study, LPS was extracted from Sabnonella paratyphi C by the hot phenol-water method,and purified with DNase I , RNase A and proteinase K. The contents of the purified polysaccharose, protein and nucleic acid of the purified LPS were detected, its bioactivity was detected with tachypleus amebocyte lysate （TAL）. The results showed that the productivity of the purified LPS was 1.48%, the proportions of the average polysaccharose, protein and nucleic acid were 3.84%, 1.49% and 5.45%. The se quence of nucleic acid was lower than 100 bp. SDS-PAGE and silver stain showed that strip size was approximately 10 to 15 ku, the agglutinate activity of TAL was 10.99 ng/mL. The chromogenic limulus amobocyte lysate assay showed that the activity of LPS was 9.82 ×10^5 EU/mg. In conclusion, the purified LPS of Salmonella paratyphi C in this study was with high purity and bioaetivity.

关 键 词：脂多糖 丙型副伤寒沙门氏菌 热酚水法 鲎试剂 显色基质法 

分 类 号：Q78[生物学—分子生物学]