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Molecular Genetics Unit 7
Terms in this set (27)
What kinds of organisms naturally produce restriction enzymes?
They are found in bacteria
Why (how is it a benefit to the
Gene cloning and genetic engineering rely on the use of enzymes that cut DNA molecules at a limited number of specific locations
Short DNA sequence that is cut out
What is a sticky end and how are they used to make new arrangements of DNA?
They are the resulting double-stranded fragments that have at least one single stranded end; these short extensions can form hydrogen-bonded base pairs with complementary sticky ends on any other DNA molecules cut with the same enzyme
The enzyme that forms covalent (phosphodiester) bonds and "glues" fragments together is called
Recombinant DNA is . . .
DNA that has been formed artificially by combining constituents from different organisms; A molecule containing DNA from two different sources
Summarize the 5 steps it takes to clone a gene
a. Isolate plasmid DNA from bacterial cells and DNA from the subjects' cells
b.Cut both DNA samples with the same restriction enzyme, one that makes a single cut within the lacZ gene and many cuts within the hummingbird DNA
c. Mix the cut plasmids and DNA fragments; some join by base pairing, add DNA ligase to seal them together
d. Mix the DNA with bacterial cells that have a mutation in their own lacZ gene. Some cells take up a recombinant plasmid or other DNA molecule by transformation
e.Plate the bacteria on agar containing ampicillin and X-gal, a molecule resembling lactose. Incubate until colonies grow
List at least one possible application of this technique
These copies can be used in sequencing the gene, in producing its encoded protein, or in basic research or other applications
Describe the technique used to identify which cell clones carry the gene of interest
Colonies with nonrecombinant plasmids will be blue, because they can hydrolyze X-gal, forming a blue product. Colonies with recombinant plasmids, in which lacZ is disrupted, will be white, because they cannot hydrolyze X-gal
Describe the process of creating complementary DNA. (Remember the reason for doing this is that eukaryotes
(like us) have noncoding introns in our genes which prokaryotes don't have. When you put a eukaryotic gene into bacteria and want that gene transcribed and protein made, bacteria don't have the means to splice out the introns, so it is done before hand by creating cDNA and putting that into the bacteria.)
A DNA molecule made in vitro using mRNA as a template and the enzyme reverse transcriptase. A cDNA molecule therefore corresponds to a gene but lacks the introns present in the DNA of the genome.
PCR stands for . . .
Polymerase chain reaction
The key ingredients for a PCR reaction are template DNA, nucleotides, single stranded DNA ___________________, and a special kind of _______________________.
What is the overall purpose of PCR?
In this technique, any specific target segment within one or many DNA molecules can be quickly amplified (copied many times) in a test tube; with automationj, PCR can make billions of copies of a target segment of DNA in a few hours, significantly faster than the days it would take to make the same number of copies by screening a DNA library for a clone with the desired gene and letting it replicate within host cells
What is gel electrophoresis?
Studying DNA molecules; This technique uses a gel made of a polymer, such as a polysaccharidel; the gel acts as a molecular sieve to seperate nucoeic acids or proteins on the basis of size, electrical charge, and other physical properties
In a gel, longer DNA molecules move faster/slower (circle one) and shorter DNA molecules move faster/slower
restriction fragment length polymorphisms?
differences in DNA cause variation in the location of restriction sites between individuals of a population. When the DNA is digested, the fragment lengths for an individual will vary compared to another and can be used to identify and distinguish one person's DNA from another's; a variation in the length of restriction fragments produced by a given restriction enzyme in a sample of DNA. Such variation is used in forensic investigations and to map hereditary disease.
describe at least four different practical uses of DNA technology
Summarize pages 403-406 in the space below
Review: In almost all cells differences between cells in a multicellular organism are due to differences in gene ____________________. This means turning genes on and off. Every cell has every gene!!!!
Describe the work done with carrots and what it means that plant cells remain totipotent.
In plants, at least, mature cells can "dedifferentiate" and then give rise to all the specialized cell types of the organism. Any cell with this potential is said to be totipotent
Describe the nuclear transplantation experiments done with frog eggs
Summarize the 6 steps used to clone Dolly, the first mammal cloned from adult donor cells.
Describe what stem cells are and why they could be so useful in terms of human health and disease.
What are homeotic genes?
What is apoptosis?
What actually causes the cells to die?
Why is this process critical in the development of all animals?
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