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Ch. 15 - Recombinant DNA Technology and Its Applications
Sanders - Genetic Analysis: An Integrated Approach 3rd Edition
Sanders3rd EditionGenetic Analysis: An Integrated ApproachISBN: 9780135564172Not the one you use?Change textbook
Chapter 15, Problem 1

What purpose do the bla and lacZ genes serve in the plasmid vector pUC18?

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1
Understand that plasmid vectors like pUC18 are used in molecular cloning to carry foreign DNA into host cells and allow for selection and identification of successful clones.
Recognize that the bla gene in pUC18 encodes the enzyme beta-lactamase, which provides resistance to the antibiotic ampicillin, allowing for selection of bacteria that have taken up the plasmid.
Know that the lacZ gene encodes the enzyme beta-galactosidase, which can cleave substrates like X-gal to produce a blue color, enabling blue-white screening to distinguish recombinant plasmids from non-recombinant ones.
Understand that insertion of foreign DNA into the multiple cloning site within the lacZ gene disrupts its function, so colonies with recombinant plasmids remain white, while those with non-recombinant plasmids turn blue on X-gal plates.
Summarize that bla allows for antibiotic selection, and lacZ enables visual screening of recombinant clones, both essential for efficient identification of successful cloning events using pUC18.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Plasmid Vectors

Plasmid vectors are circular DNA molecules used to carry foreign genes into host cells. They contain essential elements like an origin of replication and selectable markers, enabling replication and identification of transformed cells.
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bla Gene (Beta-lactamase)

The bla gene encodes beta-lactamase, an enzyme that provides resistance to ampicillin. In plasmid vectors like pUC18, it serves as a selectable marker, allowing only bacteria that have taken up the plasmid to grow in the presence of ampicillin.
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lacZ Gene and Blue-White Screening

The lacZ gene encodes beta-galactosidase, which cleaves X-gal to produce a blue color. In pUC18, disruption of lacZ by inserted DNA prevents blue color formation, enabling blue-white screening to distinguish recombinant (white) from non-recombinant (blue) colonies.
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Related Practice
Textbook Question

The results shown are from a DNA test for four genes used in a paternity identification case. DNA for the mother (M) and her child (C) are shown along with DNA from two possible fathers, F1 and F2. In the 'C' column, label the DNA bands contributed by the mother with 'M' and the DNA bands contributed by the father with 'F.'

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Textbook Question

The results shown are from a DNA test for four genes used in a paternity identification case. DNA for the mother (M) and her child (C) are shown along with DNA from two possible fathers, F1 and F2. Based on the exclusion principle, is either man excluded as the possible father? Explain.

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Textbook Question

The results shown are from a DNA test for four genes used in a paternity identification case. DNA for the mother (M) and her child (C) are shown along with DNA from two possible fathers, F1 and F2. What can you conclude based on the DNA results available?

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Textbook Question

The human genome is 3×10⁹ bp in length.

How many fragments would be predicted to result from the complete digestion of the human genome with the following enzymes: Sau3A (˘GATC), BamHI (G˘GATCC), EcoRI (G˘AATTC), and NotI (GC˘GGCCGC)?

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Textbook Question

The human genome is 3×10⁹ bp in length.

How would your initial answer change if you knew that the average GC content of the human genome was 40%?

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Textbook Question

Ligase catalyzes a reaction between the 5′ phosphate and the 3′ hydroxyl groups at the ends of DNA molecules. The enzyme calf intestinal phosphatase catalyzes the removal of the 5′5′ phosphate from DNA molecules. What would be the consequence of treating a cloning vector, before ligation, with calf intestinal phosphatase?

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