13.34. A DNA fragment containing yeast centromere DNA was cloned into a TRP ARS plasmid, YRP7, causing the plasmid to become mitotically very stable (that is, the plasmid was transmitted during mitotic divisions to each daughter cell). The assay for mitotic stability consists of growing a transformed cell without selection for the plasmid and determining the number of Trp+ colonies remaining after 20 generations of growth under conditions that are not selective for the plasmid. To identify the region of the cloned fragment that contained centromere DNA, you cut the initial fragment into smaller pieces, reclone those pieces into YRP7, and test for mitotic stability. Based on the map that follows and results of the mitotic stability assay, where is the centromeric DNA located? (On the map, B, H, and S refer to recognition sites for three different restriction enzymes; the numbers on the map are restriction fragment sizes in kb.) H B 3.5 -2.0- t-1.0–→t-0.6-† S S Results of Mitotic Stability Assay Percentage of Trp* colonies: after 20 generations Plasmid DNA YRP7 0.9 YRp7 + 5.5 kb BamHI (B) 68.1 YRP7 + 3.5 kb BamHI-HindlII (H) 0.5 YRP7 + 2.0 kb BamHI-HindlII YRP7 + 0.6 kb Sau3A (S) 80.3 76.2 YRP7 + 1.0 kb HindlII-Sau3A 0.7
13.34. A DNA fragment containing yeast centromere DNA was cloned into a TRP ARS plasmid, YRP7, causing the plasmid to become mitotically very stable (that is, the plasmid was transmitted during mitotic divisions to each daughter cell). The assay for mitotic stability consists of growing a transformed cell without selection for the plasmid and determining the number of Trp+ colonies remaining after 20 generations of growth under conditions that are not selective for the plasmid. To identify the region of the cloned fragment that contained centromere DNA, you cut the initial fragment into smaller pieces, reclone those pieces into YRP7, and test for mitotic stability. Based on the map that follows and results of the mitotic stability assay, where is the centromeric DNA located? (On the map, B, H, and S refer to recognition sites for three different restriction enzymes; the numbers on the map are restriction fragment sizes in kb.) H B 3.5 -2.0- t-1.0–→t-0.6-† S S Results of Mitotic Stability Assay Percentage of Trp* colonies: after 20 generations Plasmid DNA YRP7 0.9 YRp7 + 5.5 kb BamHI (B) 68.1 YRP7 + 3.5 kb BamHI-HindlII (H) 0.5 YRP7 + 2.0 kb BamHI-HindlII YRP7 + 0.6 kb Sau3A (S) 80.3 76.2 YRP7 + 1.0 kb HindlII-Sau3A 0.7
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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Transcribed Image Text:13.34. A DNA fragment containing yeast centromere DNA was cloned into a TRP
ARS plasmid, YRP7, causing the plasmid to become mitotically very stable (that is,
the plasmid was transmitted during mitotic divisions to each daughter cell). The
assay for mitotic stability consists of growing a transformed cell without selection
for the plasmid and determining the number of Trp+ colonies remaining after 20
generations of growth under conditions that are not selective for the plasmid. To
identify the region of the cloned fragment that contained centromere DNA, you cut
the initial fragment into smaller pieces, reclone those pieces into YRP7, and test for
mitotic stability. Based on the map that follows and results of the mitotic stability
assay, where is the centromeric DNA located? (On the map, B, H, and S refer to
recognition sites for three different restriction enzymes; the numbers on the map
are restriction fragment sizes in kb.)
H
B
3.5
-2.0-
te-1.0→t-0.6-1
S S
Results of Mitotic Stability Assay
Percentage of Trp* colonies:
after 20 generations
Plasmid DNA
YRP7
0.9
YRP7 + 5.5 kb BamHI (B)
68.1
YRP7 + 3.5 kb ВатHI-Hindil (Н)
YRP7 + 2.0 kb ВатHI-Hindil
YRP7 + 0.6 kb Sau3A (S)
0.5
80.3
76.2
YRP7 + 1.0 kb HindIII-Sau3A
0.7
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