Your friend wants to develop a new method to map the transcription start and end sites of a target gene because S1 mapping does not work well. His professor suggests starting with the cDNA synthesis method shown below. The reverse transcriptase has terminal transferase activity and adds several cytosines to the end of first strand cDNA. A template switch oligo with guanidines binds to the cytosines of cDNA and serve as new template. Your friend does not understand how a cDNA synthesis method can be used for transcript start and end sites mapping and asks you for help. LNA-containing TSO GrG+G CCC Poly(A)* RNA AAAAAAAAAA Oligo(dT) primer Reverse transcription and terminal transferase AAAAAAAAAA Q4: As a hint for transcription start site mapping, his professor further suggests using a gene specific primer for reverse transcription instead of oligo(dT) primer. Please finish the logical design of the method and briefly explain how the method works. Q5: To use the template switch mechanism for transcription end site mapping, how will you achieve target gene specificity? Please finish the logical design of the method and briefly explain how the method works.

Your friend wants to develop a new method to map the transcription start and end sites of a target gene because S1 mapping does not work well. His professor suggests starting with the cDNA synthesis method shown below. The reverse transcriptase has terminal transferase activity and adds several cytosines to the end of first strand cDNA. A template switch oligo with guanidines binds to the cytosines of cDNA and serve as new template. Your friend does not understand how a cDNA synthesis method can be used for transcript start and end sites mapping and asks you for help. LNA-containing TSO GrG+G CCC Poly(A)* RNA AAAAAAAAAA Oligo(dT) primer Reverse transcription and terminal transferase AAAAAAAAAA Q4: As a hint for transcription start site mapping, his professor further suggests using a gene specific primer for reverse transcription instead of oligo(dT) primer. Please finish the logical design of the method and briefly explain how the method works. Q5: To use the template switch mechanism for transcription end site mapping, how will you achieve target gene specificity? Please finish the logical design of the method and briefly explain how the method works.

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Description: Your friend wants to develop a new method to map the transcriptionstart and end sites of a target gene because S1 mapping does notwork well. His professor suggests starting with the cDNA synthesismethod shown below. The reverse transcriptase has ter

Biology (MindTap Course List)

11th Edition

ISBN:9781337392938

Author:Eldra Solomon, Charles Martin, Diana W. Martin, Linda R. Berg

Publisher:Eldra Solomon, Charles Martin, Diana W. Martin, Linda R. Berg

Chapter13: Gene Expression

Section: Chapter Questions

Problem 16TYU

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Below is the double stranded DNA sequence of part of a hypothetical yeast genome encoding a very small gene. Transcription starts at nucleotide immediately following the promoter. The termination sequence is TATCTC. How many amino acids will this protein have? 5' TCATGAGATA GCCATGCACTA AGGCATCTGA GTTTATATCT CA 3' 3' AGTACTCTAT CGGTACGTGAT TCCGTAGACT CAAATATAGA GT 5'
A protein has the following amino acid sequence: Met-Tyr-Asn-Val-Arg-Val-Tyr-Lys-Ala-Lys-Trp-Leu-Ile-His-Thr-Pro You wish to make a set of probes to screen a cDNA library for the sequence that encodes this protein. Your probes should be at least 18 nucleotides in length. Q. Which amino acids in the protein should be used to construct the probes so that the least degeneracy results?
Consider the Rho-dependent terminator sequence 5’CCCAGCCCGCCUAAUGAGCGGCCUUUUUUUU-3’. What affect would a point mutation at any one of the bolded and underlined nucleotides disrupt termination of transcription? Group of answer choices 1.Mutation in one of these nucleotides would disrupt base pairing, but not affect the formation of the hairpin and termination proceeds. 2.Mutation in one of these nucleotides would have no affect on base pairing, so the termination hairpin is formed and termination proceeds. 3.Mutation in one of these nucleotides would not disrupt base pairing, but would prevent the formation of the hairpin and disrupt termination. 4.Mutation in one of these nucleotides would disrupt base pairing, preventing the formation of the hairpin and disrupting termination.
The following diagram represents a transcription unit on a DNA molecule. a. Assume that this DNA molecule is from a bacterial cell. Draw the approximate locations of the promoter and terminator for this transcription unit. b. Assume that this DNA molecule is from a eukaryotic cell. Draw the approximate location of an RNA polymerase II promoter.
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Identify the open reading frame in the following DNA sequence, the protein that this gene encodes for, its function, and the source.…
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