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Make a drawing of the double recombinant plasmid containing all four fragments. Label fragment sizes, ORI (origin of replication, location of restriction sites, and location of the antibiotic resistance gene(s). number of bp for the recombinant molecule and label it on your drawing. Use Figure 15.1 for reference. Calculate the total are complementary/cut any combination of the four digested fragments is possible as long as the sticky ends same enzyme. Note that by the

Make a drawing of the double recombinant plasmid containing all four fragments. Label fragment sizes, ORI (origin of replication, location of restriction sites, and location of the antibiotic resistance gene(s). number of bp for the recombinant molecule and label it on your drawing. Use Figure 15.1 for reference. Calculate the total are complementary/cut any combination of the four digested fragments is possible as long as the sticky ends same enzyme. Note that by the

Human Anatomy & Physiology (11th Edition)
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
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Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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on which contains genes for both ampicillin resistance and kanamycin resistance. In this laboratory, you will use your digested antibiotic plasmids (Figure 15.1) from last week's laboratory to recon- struct a recombinant plasmid which possesses both the ampicillin resistance and kanamycin resistance genes. You will begin by heating your restriction digests of pAMP and 156 amp' PAMP 4539 BP ORI amp' 3755 BP Digest with BamHI and Hindill W BamHI Exploring Biology in the Laboratory will be mixed with DNA ligase plus ATP and incubated at room temperature. Complementary BamHI and HindIII "sticky ends" hydrogen-bond to align restriction fragments. Ligase catalyzes the formation of phosphodiester bonds that covalently link the DNA fragments to form stable recombinant DNA molecules. 784 BP/ 2332 BP BamHI 1875 BP Hindill kan' 12 PKAN 4207 BP ORI kan' Hindill FIGURE 15.1 Top: PAMP and PKAN plasmid maps showing locations of origins of replication (ORI) and genes for ampicillin (amp) and kanamycin (kan) resistance in black. Bottom: Restriction digests of PAMP and PKAN showing locations of restriction enzyme (BamHI and HindIII) cutting sites and resulting DNA fragments including numbers of DNA base pairs (BP).
Transcribed Image Text:on which contains genes for both ampicillin resistance and kanamycin resistance. In this laboratory, you will use your digested antibiotic plasmids (Figure 15.1) from last week's laboratory to recon- struct a recombinant plasmid which possesses both the ampicillin resistance and kanamycin resistance genes. You will begin by heating your restriction digests of pAMP and 156 amp' PAMP 4539 BP ORI amp' 3755 BP Digest with BamHI and Hindill W BamHI Exploring Biology in the Laboratory will be mixed with DNA ligase plus ATP and incubated at room temperature. Complementary BamHI and HindIII "sticky ends" hydrogen-bond to align restriction fragments. Ligase catalyzes the formation of phosphodiester bonds that covalently link the DNA fragments to form stable recombinant DNA molecules. 784 BP/ 2332 BP BamHI 1875 BP Hindill kan' 12 PKAN 4207 BP ORI kan' Hindill FIGURE 15.1 Top: PAMP and PKAN plasmid maps showing locations of origins of replication (ORI) and genes for ampicillin (amp) and kanamycin (kan) resistance in black. Bottom: Restriction digests of PAMP and PKAN showing locations of restriction enzyme (BamHI and HindIII) cutting sites and resulting DNA fragments including numbers of DNA base pairs (BP).
3 Make a drawing of the double recombinant plasmid containing all four fragments. Label fragment sizes, ORI (origin of replication, location of restriction sites, and location of the antibiotic resistance gene(s). Calculate the total number of bp for the recombinant molecule and label it on your drawing. Use Figure 15.1 for reference. Note that any combination of the four digested fragments is possible as long as the sticky ends are complementary/cut by the same enzyme. 160 amp -Bam HI site PAMP/KAN 5630bp Exploring Biology in the Laboratory Kan I -Hindill site Lorigin of replication
Transcribed Image Text:3 Make a drawing of the double recombinant plasmid containing all four fragments. Label fragment sizes, ORI (origin of replication, location of restriction sites, and location of the antibiotic resistance gene(s). Calculate the total number of bp for the recombinant molecule and label it on your drawing. Use Figure 15.1 for reference. Note that any combination of the four digested fragments is possible as long as the sticky ends are complementary/cut by the same enzyme. 160 amp -Bam HI site PAMP/KAN 5630bp Exploring Biology in the Laboratory Kan I -Hindill site Lorigin of replication
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