Question

27. Tou nave isolated the gene for a protein called nuculin (about O KDA) normally localized in the nucleus. Based on the protein sequence analysis using a computer program, you find a potential nuclear localization signal (NLS). You want to know whether this potential NLS is sufficient to bring nuculin into the nucleus. Please design experiments to address your question. You can use what you learned in Biol 203 or Biol 300.

Answer 1

Take nuculin knockout cells . Clone whole nuculin gene and fuse it with GFP .make another clone of nuculin gene with deleted NLS and fuse it with GFP and introduce it into cells .

Now observe the cells under flourescence microscope .

If cells with whole nuculin gene have nuculin in the nucleus and if cells containing deleted NLS nuculin is found to have nuculin in cytoplasm then it can be deciphered that NLS is sufficient to bring nuculin into nucleus.