J4 ›› 2010, Vol. 48 ›› Issue (05): 871-875.
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WANG Jiasi, JIN Yinghua
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Mutation(F80A)on CDK2 kinase allows the selective use of an ATP analogue N6-(2-phenylethyl)-ATP (PE-ATP), which exhibits high affinity for the altered CDK2. Using modified CDK2 and its ATP analogue enables the detection of novel CDK2 substrates in cells. Modified CDK2 (F80A)myc was cloned into pCMV vector, and transfected into HeLa cells by polyfect transfection reagent. The cells expressing mutant CDK2 were selected by the ability of resistance to G418. The stably transfected cell line was identified by immunoblotting. Four out of the six mutants showed CDK2(F80A)myc expression, and two of them got a higher expression level. The higher CDK2(F80A)myc expressing cell lines can be used for screening CDK2 substrates in cells.
Key words: stable cell line; CDK2; mutation; apoptosis
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WANG Jia-Sai, JIN Yang-Hua. Establishment of a HeLa Cell Line Stably ExpressingHuman Mutated CDK2(F80A)[J].J4, 2010, 48(05): 871-875.
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