Isolation of a fragment homologous to the rp49 constitutive gene of Drosophila in the Neotropical malaria vector Anopheles aquasalis (Diptera: Culicidae).
The constitutive ribosomal gene rp49 is frequently used as an endogenous control in Drosophila gene expression experiments. Using the degenerate primer PCR technique we have cloned a fragment homologous to this gene in Anopheles aquasalis Curry, a Neotropical vector of malaria. In addition, based on this first sequence, a new primer was designed, which allowed the isolation of fragments of rp49 in two other species, Aedes aegypti (Linnaeus) and Culex quinquefasciatus Say, suggesting that it could be used to clone fragments of this gene in a number of other mosquito species. Primers were also designed to specifically amplify rp49 cDNA fragments in An. aquasalis and Ae. aegypti, showing that rp49 could be used as a good constitutive control in gene expression studies of these and other vectorially important mosquito species.
Gentile
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Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ 21040-900, Brasil.
Lima
J B P
JB
Peixoto
A A
AA
D000330:Aedes / Q000235:genetics; D000818:Animals; D000852:Anopheles / Q000235:genetics; D003465:Culex / Q000235:genetics; D009033:Culicidae / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics / Q000302:isolation & purification*; D015870:Gene Expression; D007303:Insect Vectors / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D018076:DNA, Complementary; D029721:Drosophila Proteins; D012269:Ribosomal Proteins; C107533:RpL32 protein, Drosophila