Construction and transfection experiment of a goose circovirus infectious clone / 病毒学报
Chinese Journal of Virology
;
(6): 29-34, 2012.
Artigo
em Chinês
| WPRIM
| ID: wpr-354775
ABSTRACT
A pair of primers with BamH I restriction site were designed to amplify the complete genome of goose circovirus. Two copies of the genome were ligated in tandem and cloned into pGEM-T Easy vector to construct an infectious clone named as pGEMT-2GoCV. The pGEMT-2GoCV linearized with EcoR I was transfected to negative embryos and gosling with Lipfectamine. PCR detection verified the proliferation of GoCV in geese. Some sera of the embryo transfected group were detected to be positive at 2 and 4 weeks after hatching and one bursa was detected to be positive at 4 weeks. Some sera of the gosling transfected group were also detected to be positive at 2 weeks after transfection. Furthermore, the mark in the PCR products were identified by BamH I digestion and the GoCV in positive tissue and sera were quantitated by Real-time PCR. The results showed that the virus load in positive bursa was 1.57 x 10(6) copies/mg, the virus load in positive sera were 3.52 x 10(4)-5.92 x 10(5) copies/microL. In conclusion, the infectious DNA clone constructed with two copies of full-length GoCV genome in tandem can transfect embryo and gosling and propagate the marked goose circovirus.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Virologia
/
Transfecção
/
Circovirus
/
Reação em Cadeia da Polimerase em Tempo Real
/
Gansos
/
Genética
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Virology
Ano de publicação:
2012
Tipo de documento:
Artigo
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