Detection of genetic polymorphism among and within Echinococcus granulosus strains by heteroduplex analysis of a microsatellite from the U1 snRNA genes.
Polymerase chain reaction of a pentanucleotide microsatellite in the U1 snRNA gene complex generated a multiple band pattern due to the priming of paralogous sequences. Denaturation and slow renaturation of polymerase chain reaction products allow the formation of heteroduplex DNA that can be detected by its differential mobility in polyacrylamide gel electrophoresis. Heteroduplex analysis was used to determine if the U1 snRNA microsatellite could be a useful genetic marker in Echinococcus granulosus. A U1 snRNA microsatellite fragment from E. granulosus was isolated and characterized by Southern blot and sequencing. Four E. granulosus strains were analyzed: sheep, Tasmanian sheep, cattle, and camel strains. The former two showed polymorphism and shared three of the six patterns found for sheep strain. The cattle strain displayed two patterns, and the camel strain was monomorphic. The electrophoretic profiles were used for statistical analysis in order to determine genetic distance and the relationship among strains. Heteroduplex analysis can be helpful in genotyping E. granulosus strains and is useful in detecting polymorphism within strains.
Roratto
P A
PA
Departamento de Biologia, CCNE, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
Bartholomei-Santos
M L
ML
Gutierrez
A M
AM
Kamenetzky
L
L
Rosenzvit
M C
MC
Zaha
A
A
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002162:Camelus; D002417:Cattle; D048209:Echinococcus granulosus / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005819:Genetic Markers; D020180:Heteroduplex Analysis; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic / Q000235:genetics*; D012342:RNA, Small Nuclear / Q000235:genetics*; D012756:Sheep
D005819:Genetic Markers; D012342:RNA, Small Nuclear; C405483:U1 small nuclear RNA