Experimental study of human umbilical cord blood derived stromal cells transfected with recombinant adenoviral vector co-expressing VCAM-1 and GFP / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 598-604, 2008.
Article
de Zh
| WPRIM
| ID: wpr-267928
Bibliothèque responsable:
WPRO
ABSTRACT
This study was aimed to investigate the effect of vcam-1 gene-modified human umbilical cord blood derived stromal cells (CBDSCs) on hematopoietic regulation so as to establish the experimental foundation for further study. The target gene vcam-1 was cloned into the shuttle plasmid with the report gene GFP. The recombinant shuttle plasmid was transformed into BJ5183 bacteria to recombine with backbone vector pAdeasy-l, and the recombinant adenoviral vector ad-vcam-1-gfp was confirmed after transfection with CBDSCs. The results indicated that two fragments of about 9 kb and 2 kb were obtained after digestion of recombinant plasmid pAdTrack-vcam-1 with NotIand XhoI, and single fragment of 600 bp was obtained after amplification with PCR; two fragments of about 31 kb and 4 kb were obtained after digestion of recombinant plasmid pad-vcam-1-gfp with PacI, which suggested a successful homologous recombination. The expression of vcam-1 gene in ad-vcam-1-gfp transfected CBDSCs could be detected by immunocytochemistry, RT-PCR and fluorescent microscopy. It is concluded that the recombinant adenoviral vector ad-vcam-1-gfp has been constructed successfully, and the expression of vcam-1 is up-regulated in CBDSCs transfected by gene ad-vcam-1-gfp.
Texte intégral:
1
Indice:
WPRIM
Sujet Principal:
Protéines recombinantes
/
ARN messager
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Transfection
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Adenoviridae
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Cellules stromales
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Molécule-1 d'adhérence des cellules vasculaires
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Biologie cellulaire
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Protéines à fluorescence verte
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Sang foetal
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Vecteurs génétiques
Limites du sujet:
Humans
langue:
Zh
Texte intégral:
Journal of Experimental Hematology
Année:
2008
Type:
Article