丙型副伤寒沙门氏菌脂多糖提取纯化及活性分析

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、RNase A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100 bp,SDS-PAGE电泳和银染结果显示条带主要集中在10～15 ku范围内，与2 EU/mL鲎试剂的最小凝集浓度是10.99 ng/mL，显色基质法测定其活性为9.82×10⁵ EU/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。

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

This study was to obtain lipopolysaccharide (LPS) from Salmonella paratyphi C and analyze its activity. In this study, LPS was extracted from Salmonella paratyphi C by the hot phenol-water method,and purified with DNaseⅠ, 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 sequence 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 9.82×10⁵ EU/mg. In conclusion,the purified LPS of Salmonella paratyphi C in this study was with high purity and bioactivity.