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

作　　者：杨惠珍[1] 徐克继[1] 姜昌斌[1] 夏爱娣[1] 张建华[1] 钱宗立[1] 

机构地区：[1]上海第二医科大学寄生虫学教研室

出　　处：《中国人兽共患病杂志》1995年第3期16-18,共3页Chinese Journal of Zoonoses

基　　金：国家自然科学基金

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。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.

关 键 词：刚地弓形体 DNA 蛋白质 脂肪酸 弓形体 

分 类 号：R382.5[医药卫生—医学寄生虫学]