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Chinese Journal of Biotechnology ; (12): 223-228, 2007.
Article de Chinois | WPRIM | ID: wpr-325389

RÉSUMÉ

To generate transgenic porcine which expresses human serum albumin (HSA), the HSA gene targeting vector was constructed with HSA cDNA as the gene of interestand partial porcine serum albumin (PSA) gene as homologous arms which respectively were 7.2 kb 5' regulation sequence and 2.8 kb genomic sequence from the first intron to the fourth intron. The resistant gene neo was inserted into intron 1 and tk was ligated to the 3' end of the construct. Linearized targeting construct DNA was introduced into the fibroblast cells obtained from porcine fetus by electroporation. The positive-negative selection was performed and survival clones were screened by PCR and Southern blot. Three colonies with correct homologous recombination were obtained. Our results set a good basis for the establishment of transgenic porcine by gene target and nuclear transfer methods.


Sujet(s)
Animaux , Humains , Séquence nucléotidique , Technique de Southern , Survie cellulaire , Génétique , Cellules cultivées , Clonage moléculaire , Électroporation , Foetus , Fibroblastes , Biologie cellulaire , Métabolisme , Caryotypage , Données de séquences moléculaires , Réaction de polymérisation en chaîne , Sérumalbumine , Génétique , Suidae , Transfection , Méthodes
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