Cloning of rat serum amyloid P component gene and preparation of its recombinant adenovirus vector / 重庆医科大学学报
Journal of Chongqing Medical University
;
(12)1986.
Article
in Chinese
| WPRIM
| ID: wpr-580882
ABSTRACT
Objective:
To construct the recombinant adenovirus vector carrying rat serum amyloid P component(SAP)as preparation for later use.Methods:
Rat SAP was obtained by using RT-PCR amplification and then cloned into the shutter plasmid pAdTrace-TO4.Subsequently,this newly constructed plasmid pAdTrace-TO4-SAP was followed by homologous recombination with bone plasmid pAdEasy-1 in BJ5183.Recombinant adenovirus was obtained after being packaged in human embryonic kidney cells HEK293 by lipofectAmineTM2000 mediation.After amplification the recombined adenovirus,we determined its titer by end-point dilution assay.Results:
In recombinant shuttle plasmid and recombinant adenovirus,a fragment of 700 bp detected by PCR was identical with that included in Genbank.The titer of recombinant adenovirus reached 2.8?108 pfu/ml.Conclusion:
The recombinant adenovirus containing SAP gene was constructed successfully.This will provides a good basis for further study about the influence of SAP on lung fibrosis.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Journal of Chongqing Medical University
Year:
1986
Type:
Article
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