Title: Expression and function of the hfgl2 protein in insect cells
Abstract: Objective:To induce the expression of hfgl2 protein using baculovirus expression system in Sf9 cell.Methods:To express hfgl2 in insect cells,the full length hfgl2 gene was inserted into pENTR/D-TOPO plasmid as interest gene,then through homologous recombination of the plasmid with BaculoDirect~TM linear DNA,the recombinant baculovirus DNA containing the interest gene fragment was obtained.Sf9 cells were transfected with baculovirus DNA using Cellfectin for 4 days.After 3 rounds amplification of recombinant baculovirus,the expression of hfgl2 in Sf9 cell was detected and identified by the RT-PCR and Western blotting.Results:The recombinant hfgl2 protein was stably expressed in Sf9 cell and detected by the Distinct morphological changes of Sf9 cell,RT-PCR and Western blotting.The protein product was determined at 63kDa with capability to induce one-stage clotting assays.Conclusion:The result indicates that hfgl2 gene fragment could be expressed successfully using baculovirus expression system,providing the basic material for its functional study.Also the research took prophase exploration for developing apropos diagnostic reagents using hfgl2 as their antigens.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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