. One way to determine where inside a cell a protein(protein X) normally localizes is to generate a reportergene construct containing: (i) the gene X regulatoryregion and coding sequences, and (ii) coding sequencesfor GFP fused in frame to the 3′ end of the gene Xcoding sequences just before the stop codon. A mousecontaining such a transgene will express a hybrid protein X-GFP only in those cells in which gene X is normally expressed.a. The gene X-GFP fusion gene described could begenerated by knocking in GFP coding sequencesinstead of by random insertion of a transgene.Diagram the knockin construct you could use forthis purpose.b. What might the advantage be of the knockin strategy versus the transgene strategy?

You have synthesized an enhancerless GFP reportergene in which the jellyfish GFP cDNA is placeddownstream of a basal promoter that functions inmice. You will now fuse this enhancerless reporter tothe three types of sequences listed below (x–z).a. Which of the three types of sequences would youuse for which of the three listed purposes (i–iii)? Ineach case, explain how the particular fusion wouldaddress the particular use.Types of sequences fused to the reporter:x. random mouse genome sequencesy. known mouse kidney-specific enhancerz. fragments of genomic DNA surroundingthe transcribed part of a mouse geneUses:i. to identify a gene’s enhancer(s)ii. to express GFP tissue-specificallyiii. to identify genes expressed in neuronsb. Which of the sequences (x–z) would you fuse to aparticular mouse gene of interest in order to express the protein product of the gene ectopically,that is, in a tissue in which the gene is not usuallyexpressed? Why might you want to do this experiment in the first…

A double mutant produced by uneven crossing-over contains two single nucleotide mutations that result in frame shifts and are separated by about 20 base pairs.  The first is an insertion, while the second is a deletion.  The amino acid sequences of the wildtype and mutant polypeptide in this region of the protein are as follows: Wildtype: Lys – Lys – Tyr – His – Gln – Trp – Thr – Cys – AsnDouble Mutant: Lys – Gln – Ile – Pro – Pro – Val – Asp – Met – Asn a) What are the original and double mutant mRNA sequences.  You may find it useful to use the conventional symbols Y for pyrimidine, R for purine, N for any nucleotide, and H for A,C, or T.   2. b) Which nucleotide was inserted?   3. c) Which nucleotide was deleted?

From the list given - choose all the regulatory sequences that you think would control the expression of this eukaryotic gene  THERE are multiple answer for this that I can choose  Group of answer choices A 3' UTR B PPE sequences C 5' UTR D introns E core promoter F coding sequence G DNA-bending protein H enhancer sequence I mediator protein