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Delivery of HSF1(+) protein using HIV-1 TAT protein transduction domain
1Guangzhou Institute of Biomedicine and Health (GIBH), Chinese Academy of Sciences, 510663 Guangzhou, China.
Abstract:
HSF1 is the major transcription factor of HSPs (heat shock proteins) in response to various stresses. Wild type HSF1 (heat shock transcriptional factor 1) is normally inactive, while a constitutively active form of HSF1 (HSF1(+)) can activate downstream HSP expression in the absence of stresses. Here we generated the eukaryotic vectors that expresses HSF1(+) fusion proteins, and found that HSF1(+)-TAT fusion protein was expressed and activated HSP expression. TAT, as a trans-acting factor of HIV-1, has been demonstrated to deliver functional cargo protein into living cells. HSF1(+)-TAT fusion protein was expressed in E. coli, purified, incubated with A549 cells for 8 h, Western blot analysis and luciferase reporter assay showed that HSF1(+) fusion protein was delivered into A549 cells successfully, and the accumulation of HSF1(+)-TAT fusion protein in A549 cells up-regulated HSP70 expression.
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