Construction of a hepatic stellate cells subtracted cDNA library of differentially expressed genes in normal mice and mice with Schistosomiasis japonica / 浙江大学学报(英文版)(B辑:生物医学和生物技术)
Journal of Zhejiang University. Science. B
;
(12): 280-287, 2005.
Article
Dans Anglais
| WPRIM
| ID: wpr-249220
ABSTRACT
To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosoma japonicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5alpha. The amplified library contained more than 400 positive bacterial clone, which were then hybridized with forward and backward subtracted probes for differential screening. One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Schistosoma japonicum
/
Schistosomiase artérioveineuse
/
ARN messager
/
Séquence nucléotidique
/
Banque de gènes
/
Régulation de l'expression des gènes
/
Technique de Northern
/
ADN complémentaire
/
Hépatocytes
/
Biologie cellulaire
Limites du sujet:
Animaux
langue:
Anglais
Texte intégral:
Journal of Zhejiang University. Science. B
Année:
2005
Type:
Article
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