Which pair of restriction enzymes will you use to insert EGFP gene from Add Vector 174028 in Addgene Vector 81061 between EF1-alpha-intron A and P promoter? BsrGl and BamHI EcoRI and BsrGl EcoRI and Kpnl Kpnl and BstBI

Which pair of restriction enzymes will you use to insert EGFP gene from Add Vector 174028 in Addgene Vector 81061 between EF1-alpha-intron A and P promoter? BsrGl and BamHI EcoRI and BsrGl EcoRI and Kpnl Kpnl and BstBI

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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A partial diploid in E. coli is created so that LacI is no longer expressed from the genome and is instead expressed from a plasmid. When this E. coli strain is grown in the presence of lactose and glucose, we should expect that the lac operon will be: Question 14 options: Constitutively expressed due to LacI being expressed from a plasmid Induced due to functional Catabolite Activator Protein (CAP) Repressed due to functional LacI Induced due to non-functional LacI Expressed at low levels due to non-functional Catabolite Activator Protein (CAP)
Please determine the relative location of the three restriction enzymes. Enzyme Number of fragments Size (kb) Xbal 2 17, 21 Xhol 2. 5, 33 Xbal/Xhol 5, 12, 21 Kpnl 6, 7, 58 Xbal/Kpnl 6, 7, 8, 17
The linear dsDNA genome of λ binds on the LamB receptor of E. Coli and conducts a normal lysogenic cycle. Exposure to stress will cause the excision of λ prophage from the E. Coli genome. The excised λ genome is then replicated, packaged, and released from the cell as mature λ phage particles and ready to infect other bacterial cells. Among λ phage particles,the transducing phage mediates a specific type of recombination. Understand this scenario and answer the following questions. 1. What are the basic requirements for the insertion of λ into the E. Coli genome? 2. What special features are found in the λ insertion site? 3. What type of recombination occurs with λ insertion in the E. Coli genome? 4. How you will differentiate λ transducing phage from normal λ phage? 5. What exclusive mechanism λ phage utilizes for recombination?
Arrange the steps that would be used in a laboratory to engineer a bacterium that could express the human gene coding for factor VIII. Not all steps will be placed. Isolate the human gene that produces factor VIII. Isolate the mRNA of the factor VIII gene. Generate cDNA of the factor VIII gene using reverse transcriptase. Incorrect Insert the factor VIII cDNA into a bacterial vector near a promoter site. Transform the vector into an E. coli bacterium. Human DNA is introduced into the bacterial cell using direct injection by a small needle. E. coli expresses factor VIII. Answer Bank The factor VIII protein is isolated from human tissues.
Table 1 shows a list of restriction endonucleases with their recognition sequence and the sites of cleavage indicated by arrows. Table 1 Enzyme name Recognition sequence and position of cut 5'GIAATTC3 5'G!GATCC3' 5'GIGTACC3 5'GCIGGCCGC3' 5'IGATC3' 5'GGTACIC3' 5'ALGATCT3 EcoRI ВатHI Аcс651 Notl Sau3A Kpnl BglII (i) Which restriction enzyme(s) produce blunt ends? (ii) Are there any pair of neoschizomers in the list? Explain. (iii) Are there any pair of isocaudomers in the list? Explain.
Which of the following single-stranded DNA molecules may possibly be a restriction enzyme target cut site in the double-stranded state? O ATGCCGTA GTATCTAT OATGCTACC OGTCATGAC
In this western blot, the levels of actin increase with increasing amounts/expression of the viral protein (VP). Figure description: Increasing amounts of a plasmid expressing the viral protein (0.5, 1, or 2ug) were cotransfected with TBK1 expression plasmid. Cells were harvested 24 h post-transfection and analyzed for phosphorylated TBK1 (anti-PTBK1 Ser172), total TBK1 (anti-TBK1), B-actin (anti-ß-actin), and viral protein (anti-VP) expression by Western blot analysis. VP PTBK1 (S172) TBK1 actin Virus proteins O True O False
Explain the process of how X-gal screening works with pUC19, you may build a model with boxes and arrows. 2. You are utilizing BamHI (GGATCC) restriction site and HindIII (AAGCTT) restriction site. Within pUC19, BamHI is at position 263, while HindIII is at position 233. (Hint: position is like coordinate on a map). If you manage to insert GTF2H5 in pUC19 vector, what are the sizes of fragments if you digest the pUC19-GTF2H5 (this is after insertion) with the following restriction enzyme combination after gel electrophoresis: BamHI and HindIII There is an NdeI site right in the middle of the GTF2H5 that has been inserted in pUC19, what would be the fragment sizes, if you digest with BamHI and NdeI. HindIII and NdeI BamHI, HindIII, and NdeI. 3. Design an experiment to confirm the presence of insert GTF2H5 in pUC19 vector using the following method (besides restriction analysis above), assuming that you know the sequence of GTF2H5: Southern Blot Polymerase Chain Reaction
Restriction endonucleases are bacterial enzymes that cleave duplex (double-stranded) DNA at specific nucleotide sequences. The mode of replication of the animal virus SV40 has been investigated by using restriction endonucleases that cleave SV40 DNA into a number of unique segments. Like most viruses, SV40 DNA is circular. The map positions of the 11 fragments produced by a pair of restriction endonucleases are shown on the next page. Immediately following a 5 or 10 minute pulse of radioactively labeled thymidine, labeled SV40 molecules that have completed replication during the pulse are isolated. These newly replicated DNA molecules are digested by the restriction endonucleases and the resulting fragments are analyzed for the relative amounts of pulse label they contain. The results are in the table below. Assume that at the time the label was added there was a random population of replicating SV40 DNA molecules in all possible stages of synthesis. From the information given below,…
In the following "gene library" cloning experiment Digested genomic DNA AmpR gene TCR gene TCR is tetracycline resistant marker, AmpR is ampicillin resistant marker and BamHI is the unique restriction enzyme on plasmid. A PhD student digests/cuts the plasmids with BamHI restriction enzyme and the genomic DNA with EcoRI restriction enzyme. After performing the cloning experiment and obtaining colonies on a selection plate, the obtained cells will be ..... (Hint: this question is even more challenging; the PhD student was later demoted to an MSc student). a) resistant to ampicillin and tetracycline b) sensitive to tetracycline and ampicillin c) resistant to tetracycline and sensitive to ampicillin d) resistant to ampicillin and sensitive to tetracycline e) sensitive to ampicillin and tetracycline BamHI
Describe the fate of the λ phage during the infection process with mutants in the following genes: CI CII CIII N Cro att Q Describe the fate of the λ phage during the infection process with mutants in the following areas: OR1 OR3 PL PR PRE PRM tL1 tR1
Describe how P1vir transduction can be used to introduce a gene mutation into E. coli USING DIAGRAMS ONLY. small figures to explain the diagrams is allowed. You should describe / show both the molecular biology and the transduction procedure