Biological Science (7th Edition)
7th Edition
ISBN: 9780134678320
Author: Scott Freeman, Kim Quillin, Lizabeth Allison, Michael Black, Greg Podgorski, Emily Taylor, Jeff Carmichael
Publisher: PEARSON
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Chapter 20, Problem 12PIAT
Transgenic plants usually contain genes of bacterial plasmid origin. In a recent study, researchers designed a strategy that avoided using any plasmid genes. They transformed cells from a susceptible potato variety with a potato blight resistance gene cloned from a resistant variety. Next, to determine which plants from this group were also free of plasmid DNA (cloning vector) sequences, they performed PCR using primers specific for the plasmid. The positive control lane shows PCR amplification of plasmid DNA only, and the negative control lane shows an attempted PCR amplification of no added DNA. Based on the gel analysis of PCR products shown below, which plants only contain the potato gene? Explain your answer (See BioSkills 10 and BioSkills 6 for an overview of PCR and for help in reading a gel).
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Scientists modified the tumor-inducing (Ti) plasmid, a naturally occurring plant plasmid found in Agrobacterium tumefaciens, to create a tool used to introduce any gene of interest into plant cells. They created a binary system because a single Ti plasmid is too large to be easily manipulated. One part of the system is a disarmed
plasmid, and the second part is a transformation vector. The following sentences describe the function of key DNA elements in the system.
Virulence
region
Genes for
conjugative transfer
Disarmed
Ti plasmid
(T-region removed)
ori
Kan selectable marker
plant selectable marker
constitutive promoter
T-region
conjugative transfer
virulence
Kan
(bacterial
selectable
marker)
Amp selectable marker
Plant selectable marker
(e.g., herbicide resistance)
Constitutive
promoter
T-DNA left
border
Place the terms in the appropriate blanks to complete the sentences. Not all terms will be used.
3' transcriptional terminator
MCS (inserted
gene of interest)
T-region
Transformation…
You are cloning a gene called ice, which will help strawberry plants
survive in cold weather, into a test group of strawberry plants. Select
the events you would use to genetically engineer these plants.
Insert the ice gene into the T-DNA region of a Ti plasmid;
transform Agrobacterium tumefaciens with this plasmid; Inoculate a
culture of strawberry plant cells with Agrobacterium tumefaciens; Select
plant cells that have taken up the ice gene.
Insert the ice gene into the T-DNA region of the Agrobacterium
tumefaciens chromosome; Inoculate a culture of strawberry plant cells
with Agrobacterium tumefaciens; Select plant cells that have taken up
the ice gene.
O Insert the ice gene into a Ti transposon; Introduce the transposon
into Agrobacterium tumefaciens Ti plasmid: Inoculate a culture of
strawberry plant cells with Agrobacterium tumefaciens: Select plant cells
that have taken up the ice gene.
Insert the ice gene into a Ti bacteriophage; transduce Agrobacterium
tumefaciens with this…
When using a conventional plasmid cloning vector containing a b-galactosidase gene, it is possible to perform a "blue-white screen" to determine which bacteria have taken up a plasmid into which a DNA fragment as been inserted, as opposed to those that have taken up just reclosed plasmid vector, by growing the transformed cells on nutrient agar plates containing the artificial b-gal substrate X-gal. Will bacteria that have taken up a plasmid into which a DNA fragment has been inserted form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.
Chapter 20 Solutions
Biological Science (7th Edition)
Ch. 20 - Prob. 1TYKCh. 20 - What is a plasmid? a. an organelle found in many...Ch. 20 - CAUTION The human genome size is 3 billion base...Ch. 20 - Prob. 11PIATCh. 20 - Transgenic plants usually contain genes of...Ch. 20 - Prob. 13PIATCh. 20 - 14. PROCESS OF SCIENCE Why was it important to...Ch. 20 - Prob. 15PIATCh. 20 - Prob. 16PIAT
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