A plasmid, pUC18, contains the ampicillin-resistance gene, the origin of replication, and the B- gal gene, which codes for the B-galactosidase protein. This protein can break down the synthetic chemical X-gal, producing a blue product that stains the entire cell blue (but is harmless to the bacteria). At the beginning of the ß-gal gene there are several unique restriction sites (some of them are shown in the diagram below). You wish to clone a 1.0-kb Xbal fragment into the pUC18 plasmid, so you cut the plasmid with Xbal and, after removing the enzyme, mix the Xbal-cut plasmid with the 1.0-kb fragment, ligate, and transform competent bacteria. Pati EcoRI Xbal- B-gal R Amp ori Figure: pUC18 plasmid map (a) On what medium would you grow your transformed bacteria? (b) Do you expect the bacteria carrying plasmid pUC18 (without the insert) to be blue or white when grown in the presence of X-gal? Explain.
A plasmid, pUC18, contains the ampicillin-resistance gene, the origin of replication, and the B- gal gene, which codes for the B-galactosidase protein. This protein can break down the synthetic chemical X-gal, producing a blue product that stains the entire cell blue (but is harmless to the bacteria). At the beginning of the ß-gal gene there are several unique restriction sites (some of them are shown in the diagram below). You wish to clone a 1.0-kb Xbal fragment into the pUC18 plasmid, so you cut the plasmid with Xbal and, after removing the enzyme, mix the Xbal-cut plasmid with the 1.0-kb fragment, ligate, and transform competent bacteria. Pati EcoRI Xbal- B-gal R Amp ori Figure: pUC18 plasmid map (a) On what medium would you grow your transformed bacteria? (b) Do you expect the bacteria carrying plasmid pUC18 (without the insert) to be blue or white when grown in the presence of X-gal? Explain.
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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Bacterial Genomics
The study of the morphological, physiological, and evolutionary aspects of the bacterial genome is referred to as bacterial genomics. This subdisciplinary field aids in understanding how genes are assembled into genomes. Further, bacterial or microbial genomics has helped researchers in understanding the pathogenicity of bacteria and other microbes.
Transformation Experiment in Bacteria
In the discovery of genetic material, the experiment conducted by Frederick Griffith on Streptococcus pneumonia proved to be a stepping stone.
Plasmids and Vectors
The DNA molecule that exists in a circular shape and is smaller in size which is capable of its replication is called Plasmids. In other words, it is called extra-chromosomal plasmid DNA. Vectors are the molecule which is capable of carrying genetic material which can be transferred into another cell and further carry out replication and expression. Plasmids can act as vectors.
Question
Transcribed Image Text:A plasmid, pUC18, contains the ampicillin-resistance gene, the origin of replication, and the ß -
gal gene, which codes for the B-galactosidase protein. This protein can break down the synthetic
chemical X-gal, producing a blue product that stains the entire cell blue (but is harmless to the
bacteria). At the beginning of the B-gal gene there are several unique restriction sites (some of
them are shown in the diagram below). You wish to clone a 1.0-kb Xbal fragment into the pUC18
plasmid, so you cut the plasmid with Xbal and, after removing the enzyme, mix the Xbal-cut
plasmid with the 1.0-kb fragment, ligate, and transform competent bacteria.
Pati
Xbal
EcoRI
B-gal
A
Amp
ori
Figure: pUC18 plasmid map
(a) On what medium would you grow your transformed bacteria?
(b) Do you expect the bacteria carrying plasmid pUC18 (without the insert) to be blue or white
when grown in the presence of X-gal? Explain.
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