17 Questions
What is the purpose of electroporation in the formation of transgenic microorganisms?
To increase the permeability of bacterial membranes
How do gene markers help in identifying bacteria that have taken up the DNA?
By inactivating if DNA is not taken up
Which technique is commonly used in DNA profiling to identify individuals based on their DNA characteristics?
Polymerase chain reaction (PCR)
What is the role of antibiotic restraint genes as gene markers in transgenic microorganisms?
To kill bacteria that take up DNA
How does PCR facilitate gene technologies such as DNA profiling?
By amplifying DNA samples
Why are fluorescent markers incorporated into plasmids in genetic studies?
To make bacteria glow under UV light
What is the significance of comparing genomes between species?
Determining evolutionary relationships
Which enzyme catalyzes the formation of a double strand of DNA from a single strand of RNA?
Reverse transcriptase
What does gene sequencing allow for in terms of predicting sequences?
Predicting amino acid sequences in polypeptides
What is the main goal of the Human Genome Project?
Determining the sequence of bases in a human genome
What is a potential application of the Human Genome Project according to the text?
Screening for mutated sequences
What are restriction endonucleases?
Enzymes that cut DNA at specific sequences
Why are staggered cuts by restriction endonucleases more useful than blunt cuts?
They leave sticky ends on the DNA
What is the role of DNA ligase in gene cloning?
It joins sticky ends of DNA fragments
Why are sticky ends important in gene cloning?
They allow for easier cloning of DNA fragments
What would happen if a DNA fragment was placed directly into a cell without using a vector?
The DNA fragment would be digested by enzymes
What is the purpose of using the same restriction enzyme to cut both the plasmid and the gene during gene cloning?
To generate complementary ends for joining
Study Notes
Formation of Transgenic Microorganisms
- Electroporation is used to stimulate bacterial cells to take up plasmids, increasing the permeability of bacterial membranes.
- The process involves using calcium salts and rapid temperature change from 0 to 40 degrees.
- Gene markers are used to check whether the DNA has been taken up by the bacteria.
- Types of gene markers include antibiotic restraint genes, fluorescent markers, and enzyme markers.
Gene Technologies
- DNA profiling is a forensic technique used to identify individuals by characteristics of their DNA.
- DNA profiling can also be used to determine genetic relationships between organisms.
- Polymerase chain reaction (PCR) is used to amplify DNA by making millions of copies of a given DNA sample.
- Comparing genomes between species allows evolutionary relationships between species to be determined.
- Comparing genomes of individuals enables differences to be identified, which can be used for development of personalized medicine.
Gene Sequencing
- The Human Genome Project has successfully determined the sequence of bases of a human genome.
- Potential applications of gene sequencing include screening for mutated sequences, carriers, and pre-implantation screening.
- Gene sequencing also allows for the prediction of amino acid sequences in polypeptides and the development of synthetic biology.
Recombinant DNA Technology
- Recombinant DNA technology involves manipulating DNA using various processes.
- Reverse transcriptase can be used to make DNA from a single strand of RNA.
- Restriction endonucleases are enzymes that cut DNA at specific sequences, usually six base pairs in length.
- Staggered cuts made by restriction endonucleases leave sticky ends on the DNA, which can be used to attach DNA fragments together.
In-Vivo Gene Cloning
- Vectors are used to insert DNA into cells, as DNA fragments would be digested by enzymes if inserted directly.
- Plasmids from bacterial cells are naturally occurring vectors.
- Isolated DNA fragments can be placed in plasmids using restriction enzymes to create complementary ends.
- DNA ligase is used to form phosphodiester linkages, sealing the DNA fragments together.
Test your knowledge on the creation of recombinant DNA molecules and the use of electroporation in stimulating bacterial cells to take up plasmids. Understand how electroporation increases the permeability of bacterial membranes to enhance success.
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