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弓形虫人源、畜源分离株的生化特性研究

Biochemical Characterization on Toxoplasma gondii of Human &Animal Origins

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【作者】 杨惠珍徐克继姜昌斌夏爱娣张建华钱宗立

【Author】 Yang Huizhen; ZongLi Qian et al(Department of Parasitology,Shanghai Second Medical University,Shanghai,200025)

【机构】 上海第二医科大学寄生虫学教研室

【摘要】 本文采用弓形虫特异DNA探针杂交技术,聚丙烯酰胺电泳,免疫印迹试验,色质谱分析对弓形虫人源分离的SH、Zs2株,猪源分离的CN,PP株,兔源RT株等11个分离株在核酸、蛋白质、脂肪酸方面与国际标准人株RH株进行分子生化特性的比较与分析,实验结果表明用32P标记弓形虫DNA探针(1.1Kb)能与各虫株DNA杂交,在2.0Kb位点上显示出明显的杂交带;SDS-PAGE电泳蛋白质分子量范围在1-120KD间,以14、18、22、30、35、37、41、43、52、76KD为各虫株共有的条带,Westernblot分析,各虫株在30、52、76KD均能被抗弓形虫IgG识别;有机质谱仪检测脂肪酸组分中各虫株均有6种主要脂肪酸组成(c14一C18)分别为肉豆蔻酸、棕榈油酸、软脂酸、硬脂酸、油酸和亚油酸,其中油酸和软脂酸占总脂肪酸含量的70%,各虫株相对含量有所差异。实验结果为弓形虫分离株的生化特性和鉴定工作提供了科学理论依据。

【Abstract】 Eleven isolates of Toxoplasma gondii,respectively coded as SHn,ZH2 of human origin,CN,PP of swine origin and RT of rabbit origin,were subjected to biochemical characterization for nucleic acid,protein and fatty acid analysis by means of Toxo-Specifc DNA probing,PAGE electrophoresis,Western blotting and GC/MS techniques with a parallel comparison to the international standard RH strain of humans. Results indicated that the 32P labelled probe of a 1.1 kb Toxoplasma-fragment could successively hybridize to DNA of alI the various isolates demonstrating a distinct hybridizing band at the site of 2.0 kb;SDS-PAGE separation of the parasite proteins revealed a molecular weight spectrum ranging from 1-120 KD and the bands at 14, 18,22, 30, 35, 37, 41, 43, 52, 76 KD were shared in all tested isolates. Western blotting analysis showed a common recognition of the bands 30,52 and 76 KD in all isolatesby a Toxo-specific IgG antibodies. Six main fatty acid(MFA) invoived in C14-C18,demonstrated in relative percentage ratio,which comprised palmitoleic acid, oleic acid,stearic acid,nutmeg acid,biorleic acid and palmitic acid,with the first two for an average of 70%(52-84%).Results of this study thus provided biochemical information which may serve as criteria for biological identification of the Toxoplasma isolates.

【基金】 国家自然科学基金
  • 【文献出处】 中国人兽共患病杂志 ,CHINESE JOURNAL OF ZOONOSES , 编辑部邮箱 ,1995年03期
  • 【分类号】R382.5
  • 【被引频次】11
  • 【下载频次】79
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