Title: Cloning of rat serum amyloid P component gene and preparation of its recombinant adenovirus vector
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.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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