RESUMO
A new method has been developed allowing the identification and relative quantification of different forms of mRNA after RNA editing. This method was applied to the serotonin 2c receptor mRNA that potentially exhibits 32 different forms after adenosine to inosine editing at five different sites located in a row of 13 nucleotides. CE was used to characterize fluorescently labeled ssDNA molecules on the basis of their conformational polymorphism. The relative amount of these 32 mRNA forms has been estimated by measuring the fluorescence intensity of each individual DNA strand. Accuracy of quantification was established by diluting one form into another or into a mixture of cDNA, showing linear and precise proportion of each form (0.06 Assuntos
Eletroforese Capilar/métodos
, Edição de RNA
, RNA Mensageiro/genética
, RNA Mensageiro/metabolismo
, Receptor 5-HT2C de Serotonina/genética
, Adenosina/genética
, Animais
, Sequência de Bases
, Encéfalo/metabolismo
, Primers do DNA/genética
, DNA Complementar/genética
, DNA Complementar/isolamento & purificação
, Inosina/genética
, Camundongos
, Camundongos Endogâmicos BALB C
, Ratos
, Ratos Wistar