摘要
本文首次对分离自中国、美国、柬埔寨和澳大利亚７株贾第虫基因组ＤＮＡ，用６种限制性内切酶消化并进行琼脂糖电泳，分析比较它们基因组ＤＮＡ酶切片段多态性；同时用生物素标记的贾第虫全基因组ＤＮＡ探针对国内４株贾第虫ＤＮＡ进行Ｓｏｕｔｈｅｒｎ印迹杂交。结果表明，７株贾第虫ＤＮＡ经ＨｉｎｄⅢ和ＢｇｌⅡ酶切后，可将国内４株和国外３株贾第虫区别开；国内４株贾第虫ＨｉｎｄⅢ、ＢｇｌⅢ、ＨａｅⅢ、ＰｓｔⅠ和Ｈｉｎｆ５种内切酶酶切图谱一致；而经ＡｌｕＩ酶切后，Ｃ１和ＣＲ的ＤＮＡ图谱一致，Ｃ２和Ｃ３的也一致；Ｓｏｕｔｈｅｒｎ印迹杂交显示，国内４株间的ＤＮＡ杂交带相似，提示分离自人和兔的贾第虫无严格的宿主专一性，二者可产生交叉感染；国内３株与国外２株人贾第虫ＤＮＡ图谱存在差异，其原因似与地理隔离有关。
The Restriction Fragment Length Polymorphism (RFLP)of 7 Giardia lamblia isolates, of which four (C1, C2, C3 and CR) were from China, one (PB) from America, one (CAM) from Cambodia and the other (A) from Arstralia, was analysed. Isolates C1, C2, C3, CAM and A were isloated from human being, CR and PB from rabbit and beaver respectively. Trophozoites of these isolates were cultivated axenically in modified TYI-S-33 medium. Total genomic DNA of each isolate were extracted with CTAB method and firstly digested separatly with Hind Ⅲ and Bgl Ⅱ. The 4 domestic isolates were, in addition, digested separatly with Pst Ⅰ,HaeⅢ, Hinf and Alu Ⅰ.All the DNA digested with enzymes were then carried on agarose gel electrophoresis for RFLP analysis. Besides, DNA from 4 domestic isolates degisted with AluⅠwere hybridized with a total genomic DNA probe, which were prepared from C2 and labelled with biotin,in Southern bloting.It was shown that after being digested with Hind Ⅲ and Bgl Ⅱ, the DNA banding patterns of Cl, C2,C3 and CR were identical, those of CAM, A and PB were different.In the digestion tests of PstⅠ,HaeⅢand Hinf, the DNA banding patterns obtained from 4 demostic isolates were similar.When digested with AluⅠ, the DNA banding patterns of C1 and C R were identical. Those of C2 and C3 were also identical. The Southern bloting hybridization showed that C1 and CR had identical hybridization bands, C2 and C3 identical bands too
出处
《寄生虫与医学昆虫学报》
CAS
1994年第1期1-7,共7页
Acta Parasitologica et Medica Entomologica Sinica
关键词
贾第虫
限制性内切酶
DNA
RFLP
Giardia lamblia Genomic DNA Restriction fragment Length polymorphism Southerm bloting
