Method for Determining Transduction Efficiency and Virus Dosage of Baculovirus
First Claim
1. A method for determining transduction efficiency of a baculovirus, comprising:
- providing a recombinant baculovirus, wherein the recombinant baculovirus comprises an inducible promoter and a reporter gene positioned downstream the inducible promoter;
adding the recombinant baculovirus into an incubating environment of a mammalian cell for transduction;
adding an inducer to promote expression of the reporter gene in the mammalian cell; and
analyzing a percentage of the mammalian cell expressing the reporter gene to determine a transduction efficiency of the recombinant baculovirus.
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Abstract
A method for determining transduction efficiency of baculovirus includes: providing a recombinant baculovirus, in which the recombinant baculovirus includes an inducible promoter and a reporter gene positioned downstream the inducible promoter; adding the recombinant baculovirus to incubating environment of a mammalian cell for transduction; adding an inducer to promote expression of the reporter gene in the mammalian cell; and analyzing the percentage of the mammalian cell expressing the reporter gene to determine the transduction efficiency of the recombinant baculovirus. The method provides the ability to quantitatively analyze baculovirus transduction and is a simple and faster quantitative method applicable in transduction and other research study by including an inducible promoter and a reporter gene and thus prevents from imposing excessive metabolic burden to the cells. A method for determining virus dosage of baculovirus applied in genetic therapy is also disclosed.
3 Citations
19 Claims
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1. A method for determining transduction efficiency of a baculovirus, comprising:
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providing a recombinant baculovirus, wherein the recombinant baculovirus comprises an inducible promoter and a reporter gene positioned downstream the inducible promoter; adding the recombinant baculovirus into an incubating environment of a mammalian cell for transduction; adding an inducer to promote expression of the reporter gene in the mammalian cell; and analyzing a percentage of the mammalian cell expressing the reporter gene to determine a transduction efficiency of the recombinant baculovirus. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for determining virus dosage of a baculovirus, comprising:
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providing a recombinant baculovirus, wherein the recombinant baculovirus comprises an inducible promoter and a reporter gene positioned downstream the inducible promoter; adding the recombinant baculovirus into an incubating environment of a mammalian cell for transduction; adding an inducer to promote expression of the reporter gene in the mammalian cell; analyzing a percentage of the mammalian cell expressing the reporter gene to determine a transduction efficiency of the recombinant baculovirus; and determining a virus dosage of the recombinant baculovirus based on the transduction efficiency of the recombinant baculovirus. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification