Chemical Incorporation of Fluorescent Probe FlAsH on the Hha Protein: Application to the Study of Complex Hha/H-NS The objective of this investigation was the development of studies of incorporation of the probe F1AsH as a possible tool for the study in vitro and in vivo of the protein Hha and the interaction with H-NS. A protein was constructed with the specific sequence “CCPGCC” able to join the fluorescent molecular probe FLAsH; later, processes of transformation, expression, and purification were developed, with which it was demonstrated that even with the introduction of an additional no native sequence in the protein, it could be obtain protein in a good amount, stability, yield and whit a high level of purity. The optimization of the protocol of incorporation of FLAsH was made considering the yield and the time of reaction. FLAsH was characterized with fluorescence and was verified with MALDI TOF. The incorporation HhaCCPGCC1/ FLAsH could not be obtained in a high percent as it was expected; therefore, the studies of interaction between the protein complex Hha and H-NS could not be done.
keywords:
fluorescente anisotropy, FLAsH, Hha, H-NS, molecular probe, nucleoid-associated proteins.
