GENETICS(LL)-W/CONNECT >CUSTOM<
GENETICS(LL)-W/CONNECT >CUSTOM<
6th Edition
ISBN: 9781260571561
Author: HARTWELL
Publisher: MCG CUSTOM
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Chapter 15, Problem 9P

An example of a gene-targeting DNA plasmid vector for insertion of a transgene into chloroplast DNA by biolistic transformation is shown in the following diagram. The plasmid DNA can be prepared in large quantities in E. coli before being shot into plant cells with a gene gun. Match the component of the construct with its function. (RE1, RE2, and RE3 are different restriction enzyme recognition sites unique to the plasmid vector.)

 Chapter 15, Problem 9P, An example of a gene-targeting DNA plasmid vector for insertion of a transgene into chloroplast DNA

a. spectinomycin 1. DNA that mediates resistance gene integration
b. cpDNA sequences 2. gene used to select chloroplast transformants
c. RE2 3. sequence for plasmid replication in E. coli
d. ori 4. sites at which targeting chloroplast DNA can be inserted into the vector
e. RE1 and RE3 5. site at which transgene can be inserted into the vector
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Students have asked these similar questions
A shuttle vector is a vector (usually a plasmid) constructed so that it can propagate in two different host species. One of the most common types of shuttle vectors is the yeast shuttle vector. Examples of such vectors derived from yeast are Yeast Episomal Plasmid (YEP), Yeast Integrating Plasmid (YIP) and Yeast Replicating Plasmid (YRP). Why is YEP preferred over YIP and YRP? Give your thoughts on this.
When cloning a foreign DNA fragment into a plasmid, it is often useful to insert the fragment at a site that interrupts a selectable marker(such as the tetracycline-resistance gene of pBR322). The loss of function of the interrupted gene can be used to identify clones containing recombinant plasmids with foreign DNA. With a yeast artificial chromosome (YAC) vector, it is not necessary to do this; the researcher can still distinguish vectors that incorporate large foreign DNA fragments from those that do not. How are these recombinant vectors identified?
After transformation you were asked to grow bacterial cells transformed with plasmid on a plate that had X-gal and ampicillin. X-Gal is often used as in indicator dye, which turns blue when metabolized by B-galactosidase protein and used to test if cloning experiments have worked. [Note look at the vector diagrams carefully] Briefly explain how you would find the bacterial cells that are transformed with the plasmid with the YFG inserted.

Chapter 15 Solutions

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