AGC AGA AGG GCA CGA GCC CGC UUA UUG CUA AUA CUC 1 of 5 GUA UAA GGA GGC CCA UCA A

Question

AGC AGA AGG GCA CGA GCC CGC UUA UUG CUA AUA CUC 1 of 5 GUA UAA GGA GGC CCA UCA ACA CCC UCC ACC GUC UAC GUG GCU CGU GAU AAU UGU GAGCAG GGU CAU AUU CUU AAG AUG UU CCU UCU ACU UGG UAU GUU GCG CGG GAC AAC UGC GAA CAA GGG CAC AUC CUG AAA UAG ARDNCEQGH?, L K M F P S T w Y V stop 1. Your friend (who is a graduate student) is interested in understanding the biological basis of sleep, and has determined the sequence of a 3 kilobase (i.e., 3000 bases long) mRNA molecule that is highly expressed in the mouse brain when mice are asleep, but not expressed when mice are awake. He has isolated a cDNA from a cDNA library. The sequence from the middle of the cDNA (about 1 kb in) is shown here 5'- .. .TCGAGCCCTATACTAAACTTGCTACATT. . .-3' 3 '- .. . AGCTCGGGATATGATTTGAACGATGTAA. . .-5' a) You want to design a probe that can hybridize to the cDNA molecule above. Circle the molecule(s) below that will hybridize to this cDNA molecule. (6 points) 5"-AAATCATATC-3 5 "-CAAGTTTAGT-3 5"-AGCTCGGGAT-3 5"-ATAAGTTTTT-3' 5 "-TCGTTCAAAT-3" 5'-??????????-3 ' b) If the scquence of the protcin from the portion of the mRNA that is encoded from this bit of cDN/A was CSKFSIGL, write the sequence of the mRNA that is produced from this sequence and show how CSKFSIGL is encoded by this mRNA. (4 points) c) If you wanted to use your probe(s) from Part la to do in situ hybridization (meaning, hybridizing to the mRNA in the cell), write the probe(s) that can be used for this below. (3 points)

Explanation / Answer

1. (a) DNA probe ( base sequences of a nucleic acid (ss DNA)) to detect the specific

nucleic acid chain by the process of hybridization.

The DNA probe for given sequence of cDNA in the figure is 5'-CTATACTAAA- 3', that will

attaches with 3'- GATATGATTT -5' sequence to hybridize the cDNA molecule.

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