E. coli bacterium Bacterial chromosome Plasmid Isolate plasmid O Recombinant DNA plasmid Copyright © 2009 Pearson Education, Inc. Recombinant bacterium Clone of cells Cut plasmid with enzyme Put plasmid into bacterium by transformation Gene of interest O Combine targeted fragment and plasmid DNA Add DNA ligase, which closes the circle with covalent bonds Gene of interest 8 Allow bacterium to reproduce Cut cell's DNA with same enzyme Isolate DNA Gene of interest 23 Cell with DNA containing gene of interest DNA

E. coli bacterium Bacterial chromosome Plasmid Isolate plasmid O Recombinant DNA plasmid Copyright © 2009 Pearson Education, Inc. Recombinant bacterium Clone of cells Cut plasmid with enzyme Put plasmid into bacterium by transformation Gene of interest O Combine targeted fragment and plasmid DNA Add DNA ligase, which closes the circle with covalent bonds Gene of interest 8 Allow bacterium to reproduce Cut cell's DNA with same enzyme Isolate DNA Gene of interest 23 Cell with DNA containing gene of interest DNA

Biology: The Unity and Diversity of Life (MindTap Course List)

14th Edition

ISBN:9781305073951

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter15: Studying And Manipulating Genomes

Section: Chapter Questions

Problem 6SQ

See similar textbooks

Similar questions

During DNA profiling, DNA nucleotides hybridized with probe can be detected througha) Electrophoresisb) Polymerase chain reactionc) Autoradiographyd) Hybridoma
A bacterium has a genome size of 4.4 Mb. If a researchercarries out shotgun DNA sequencing and sequences a totalof 19 Mb, what is the probability that a base will be leftunsequenced? What percentage of the total genome will beleft unsequenced?
You are cloning the genome of a new DNA virus into pUC18.You plate out your transformants on ampicillin plates containing X-gal and pick one blue colony and one white colony.When you check the size of the inserts in each plasmid (blueand white), you are surprised to fi nd that the plasmid fromthe blue colony contains a very small insert of approximately60 bp, while the plasmid from the white colony does notappear to contain any insert at all. Explain these results.
Entire sequence below needs to beamplified by PCR and subcloned into a plasmid vector. Which of the primersequences listed underneath is the correct reverse primer (6 marks)? Copy correctsequence into your answer. Why primer e) is not the right answer (4 marks)? 5'ATCTCTATTTAATATTTATGTCTATTTAAGCCTCATATTTAAAGACAGGGAAGAGCAGAACGGAGCCCCAGGCCTCTGTGTCCTTCCCTGCATTTCTGAGTTTCATTCTCCTGCCTGTAGCAGTGAGAAAAAGCTCCTGTCCTCCCATCCCCTGGACTGGGAGGTAGATAGGTAAATACCAAGTATTTATTACTATGACTGCTCCCCAGCCCTGGCTCTGCAATGGGCACTGGGATGAGCCGCTGTGAGCCCCTGGTCCTGAGGGTCCCCACCTGGGACCCTTGAGAGTATCAGGTCTCCCACGTGGGAGACAAGAAATCCCTGTTTAATATTTAAACAGCAGTGTTCCCCATCTGGGTCCTTGCACCCCTCACTCTGGCCTCAGCCGACTGCACAGCGGCCCCTGCATCCCCTTGGCTGTGAGGCCCCTGGACAAGCAGAGGTGGCCAGAGCTGGGAGGCATGGCCCTGGGGTCCCACGAATTTGCTGGGGAATCTCGTTTTTCTTCTTAAGACTTTTGGGACATGGTTTGACTCCCGAACATCACCGACGCGTCTCCTGCTG 3'a) 5' TTCCGGAAGAAGCTTATACGG 3'b) 5' CTGTGTTCACCTAATATTCCT 3'c) 5' CAGCAGGAGACGCGTCGGTGA 3'd) 5' AGGAATATTAGTATAATCCAC 3'e) 5' GACGCGTCGGTGATGTTCGGG 3’f) 5'…
#2 EcoRI --- 5’ G ↓AATTC 3’ 5’ ACG ACGTATTAGAATTCTTA TCCGCCGCCGGAATTCT CATCA 3’ 3’ TGC TGCATAATCTTAAGAATAGGCGGCGGCCTTAAGAGTAGT 5’ Restriction enzyme: Recognition sequence: Number of pieces of DNA: Type of cut:
Please write down the DNA sequence inferred from the below DNA gel. Shown are the products of a dideoxy sequencing reaction.
As you learned in this chapter,restriction enzymes are sophisticated“scissors” that geneticistsand molecular biologists routinelyuse to cut DNA for recombinant DNAexperiments. A wide variety of onlinetools assist scientists working with restrictionenzymes and manipulating recombinantDNA for different applications. Herewe explore Webcutter and Primer3, twosites that make recombinant DNA experimentsmuch easier.Exercise I – Creating a RestrictionMap in WebcutterSuppose you had cloned and sequenceda gene and you wanted to design a probeapproximately 600 bp long that could beused to analyze expression of this gene indifferent human tissues by Northern blotanalysis. Internet sites such as Webcuttermake it relatively easy to design experimentsfor manipulating recombinant DNA.In this exercise, you will use Webcutter tocreate a restriction map of human DNAwith the enzymes EcoRI, BamHI, and PstI.1. Access Webcutter at http://www.firstmarket.com/cutter/cut2.html.Go to the Study Area for…
83. A plasmid is cut with the restriction enzyme BamH1 giving fragments of 3000 and 1000 bp as identifiedby gel electrophoresis and ethidium bromide staining. In a seperate restriction digest the enzyme EcoR1gives fragments of 1000 and 1500 bp that are apparent on the agarose gel. What is the most likely size ofthe plasmid in bp? Explain why it's 4000
Genes encoding toxins are often located on plasmids.A recent outbreak has just occurred in which a bacterium that is usually nonpathogenic is producing atoxin. Plasmid DNA can be isolated from this newlypathogenic bacterial strain and separated from thechromosomal DNA. To establish whether the plasmidDNA contains a gene encoding the toxin, you coulddetermine the sequence of the entire plasmid andsearch for a sequence that looks like other toxin genespreviously identified. An easier way exists to determine whether the plasmid DNA carries the gene(s)for the toxin; this strategy does not involve DNAsequence analysis. Describe this easier method.
U have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp. What would the number of and length of the fragments be if you cut the plasmid with the following restriction enzymes or combination of enzymes? Give a schematic representation of the digestions. PscI & GsuI ______________________________________________________________ ScaI, PdmI & BsaXI ______________________________________________________________ ScaI, SspI & EheI ______________________________________________________________
You have two sewage samples that you want to check for the SARS Cov 2 viral load. Whichvariable would give you a good estimate?a. High Ct valueb. Low Ct valuec. High melting temperatured. Low melting temperaturee. None of the above The DNA fragments generated during a cycle sequencing reaction are separated by sizes usingcapillary gel electrophoresis. The determination of the actual sequence is based on detection ofa. fluorescently labelled ddNTPsb. the lengths of the fragmentsc. fluorescently labelled probesd. fluorescently labelled primers.
In a typical microbiology laboratory, reasons for no bands from a gel of a polymerase chain reaction may bedue to errors relating to omission of ingredients in the reaction mix and absence of the target sequence inthe template DNA. Based on (i) primer problem and (ii) purity/potential contamination of the target sequence, explain the reason for non-appereance on bands.