CPC C12N 15/79 (2013.01) [C12N 9/0069 (2013.01); C12N 9/12 (2013.01); C12Q 1/485 (2013.01); G01N 33/5005 (2013.01); C07K 2319/60 (2013.01); C12Y 113/12009 (2013.01)] | 3 Claims |
1. A method of determining the activity of a Rb variant in a cell, comprising the steps of:
a. providing a mammalian cell, said cell comprising a stably integrated first construct, wherein said first construct comprises a first cDNA encoding E2F1 protein which is linked to a first portion of a luciferase gene, and a second cDNA encoding Dp-1 protein;
b. preparing a linear third cDNA of a Rb variant, wherein said Rb variant contains at least one mutation as compared to wild-type Rb, and said third cDNA is linked to a second portion of a luciferase gene to form a second construct, wherein when said first construct is in close proximity with said second construct a complex is formed that emits light;
c. transfecting said second construct into said mammalian cell; and
d. determining the activity of said Rb variant by measuring any generated light emission, wherein an increase in light emission relative to a control mammalian cell is indicative of said Rb variant activity.
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