sgRNA Transgene Mapping Project Overview

Mutant #    Deletion Region    %Transcription (lungs)  %Transcription(kidneys) WT                      None                     100%                                      100%  1                          1-200                    130%                                       130% 2                       250-400                   100%                                       100% 3                       500-800                     50%                                         50% 4                      950-1100                      0%                                        100%   1) For the following gene, you notice the following results when analyzing several muntants. What type of sequence (be specific) has been mutated in mutant 1? What type of sequence (be specific) has been mutated in mutant 3? What would we call this type of mutation?  What type of sequence (be specific) has been mutated in mutant 4?

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Question: how does the flu without the track arise and what does it mean male type splicing if the flu protein isn't present in females?

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. 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?

12e which genes that are normally found in a lambda genome would be deleted in your lambda vector? briefly explain why those lambda genes are normally delete it from your lambda phage being used as cloning vectors.

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In the module, you have learned about P-element mediated transgenesis in Drosophila and the concept of using transgenes to rescue mutant phenotypes. In the figure below, you will see a wild type fly with its natural eye colour and three mutants with their eye colours changed to vermillion, white and rosy, respectively. A schematic of P-element mediated transgenesis (as shown in the lectures) is also included in the figure. Please inspect the schematic carefully and choose which of the following statements is true:    I. Injection of the white experimental transgene into the vermillion mutant embryo will not change the vermillion mutant phenotype   II. Injection of the white experimental transgene in the rosy mutant embryo will change rosy eye colour to red (wild type)   III. Injection of the white experimental transgene in the white mutant embryo will not change the white mutant phenotype   IV. Injection of the white experimental transgene in the rosy mutant…

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Pls can anyone explain how to solve this exercise and it means? thank you sm!

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. Let’s say that you have incredible skill and can isolate the white and red patches of tissue from the Drosophila eyes shown in Figure 12-24 in order to isolate mRNA from each tissue preparation. Using your knowledge of DNA techniques from Chapter 10, design an experiment that would allow you to determine whether RNA is transcribed from the white gene in the red tissue or the whitetissue or both. If you need it, you have access to radioactive white-gene DNA

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Q.7 a) You are seeking to clone a gene into pEGFP-C1 (see plasmid MCS map in Q.4). Which of the following forward primers with added RE sites (in capitals) will result in your gene being in-frame with EGFP? The start codon is underlined and the RE being used is shown in brackets. (i) 5'-nnCTCGAGatggacgatgtagcacagctt-3' (Xhol) (ii) 5'-nnGGATCCatggacgatgtagcacagctt-3' (BamHI) (iii) 5'-nnGAATTCatggacgatgtagcacagctt-3' (EcoRI) b) For the primers that will result in the gene being out-of-frame with EGFP, make alterations to the primer sequences to correct the frame.

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CA Live Remote Consider the following chart: What type of mutation is this? * DNA: TAC GCA TGG AAT MRNA: AUG CGU ACC UUA Amino acids: Met-Arg-Thr-Leu DNA: TAC GTA TGG AAT MRNA: AUG CAU ACC UUA Amino acids: Met - His - Thr- Leu substitution deletion insertion frameshift mutation

Q1A) Fig. 6.10, the blue curve suggests high level of transcription initiation for that set of samples. Why did it increase? Q1B) In the same figure, the data for the green curve was obtained with a so-called rifampicin-resistant core. You would expect that the green curve should overlap with the blue one, but it doesn’t. What is the reason?