Untitled Quiz

Recombinant DNA is formed by combining DNA segments from different sources, often different species.
This process frequently combines DNA sequences that wouldn't normally occur together.
Recombinant DNA is made in vitro, meaning it’s created in a lab setting, and placed in a single DNA molecule.
Paul Berg won a Nobel Prize in 1980 for his crucial work on the biochemistry of nucleic acids, specifically related to recombinant DNA.

Students will gain a general understanding of DNA recombination and gene cloning.
Students will understand the role of restriction endonucleases and DNA ligases.
Students will learn the key requirements for a plasmid vector.
Students will be able to outline the characteristics of host cells suitable for recombinant DNA technology.
Students will gain examples of applications using recombinant DNA technology.

cDNA is often used rather than the whole gene.
cDNA is created from mRNA, as explained in previous lectures.
The primary purpose is to create multiple copies of a gene, or its encoded protein, for various biological studies or applications.
This process typically does not involve PCR.

Basic bioscience research for analyzing protein structures and building models.
Medical advancements in developing gene therapies and vaccines.
Agricultural applications focused on plant modification, livestock (mammals and fish) improvement.
Biotechnology applications covering pharmaceuticals (antibodies, enzymes, other proteins), along with industrial chemicals.

Laboratory methods share common features, such as using bacteria and their plasmids, along with specific enzymes.
Plasmids are small, circular DNA molecules that replicate independently of the bacterial chromosome.
Cloning DNA is useful for making copies of genes and producing proteins.
Bacteria are used as host cells to create numerous copies of a gene.

Foreign DNA is inserted into a plasmid, which is then introduced into a bacterial cell.
Replication within the bacterial cell clones the plasmid, including the foreign DNA.
This process creates multiple copies of a single gene or DNA segment.
Recombinant plasmids enable the creation of tools for more complex genetic engineering applications, such as GMO manipulation.

The cloning vector (plasmid) is cut with restriction endonuclease.
The DNA fragment of interest is isolated by cleaving the chromosome with restriction endonuclease.
The fragments are then ligated to the prepared cloning vector using DNA ligase.
The recombinant vector is introduced into a host cell for replication.
The replication creates large numbers of copies of recombinant DNA.

Plasmids are also known as vectors.
Plasmids are originally small, independent bacterial DNA that can be shared and transferred (e.g., antibiotic resistance).
These molecules replicate independently of the main bacterial chromosome.
Today, plasmids are extensively modified for various research purposes (e.g., containing specialized vectors found at the Addgene website).
Plasmids used in research must have fundamental properties essential for research applications.

A multi-cloning site (MCS) is a region within a plasmid vector designed for inserting foreign DNA segments.
The MCS contains many unique restriction enzyme sites.
This allows researchers to insert genes or other DNA fragments at specific locations dictated by the choice of restriction enzyme site.
An origin of replication is a region within a plasmid molecule that is essential for autonomous replication of the plasmid in a bacterial host cell..

DNA fragments are joined using complementary ends and DNA ligase.
The result is a recombinant plasmid containing the gene(s) of interest and other necessary components.

Recombinant plasmid is introduced into a bacterial host cell via transformation process.
This procedure does not involve reproduction.

The species of E. coli are a common choice for host cells in DNA cloning.
Often, highly specialized host cells are used to produce pure/high quality recombinant DNA.
Many different strains of E. coli have been developed and are commonly used in DNA cloning, all are highly modified.

Bacteria are often chosen due to their affordability and rapid growth.
They're easy to manipulate and analyze in the lab.
They often contain a collection of modified strains, which simplifies research procedures.
The downsides include the lack of a complex RNA splicing and protein modification machinery found in eukaryotic cells – which can make them unsuitable for certain experiments.

DNA from various sources can be amplified via polymerase chain reaction (PCR). These can include plants.
Restriction enzymes are used to cut the DNA and then ligation, which links the different DNA fragments.
A separate set of enzymes are used to insert the fragments into the plasmid or vector.
This is then used to create numerous copies of the recombinant DNA.

Recombinant DNA technology is applied to drug development, animal models, genetic engineering of organisms, and industrial protein production.
These applications cover a broad spectrum of fields like medicine, agriculture, and the development of industrial products.

CRISPR-Cas9 technology simplifies and reduces costs for genome editing in eukaryotic organisms.
This method allows removing replacing genes and portions of genes – unlike the more traditional techniques.
CRISPR-Cas9 technology derived from a prokaryotic immune system against viruses.
This tool allows targeted introduction of DNA segments into organisms.
Researchers can manipulate DNA in different organisms using this technique.
The process of repairing cell DNA after a CRISPR-Cas9 cut is essential for effective genome editing.
Different mechanisms or methods can be used for this.

The goal of recombinant DNA technology is creating many copies of a specific gene (or cDNA).
The DNA is inserted into a host cell using a vector, which is cut to incorporate the desired DNA.
DNA fragment sizes are verified using gel electrophoresis.
Successful introduction of recombinant DNA can be monitored using reporter genes (e.g., antibiotic resistance).

Recombinant DNA and its manipulation are essential parts of bioscience research.
This process is furthered by specialized E. coli containing plasmids (vectors).
Modern genetic engineering is constantly improving and expanding to encompass new methods and applications.
Recombinant DNA technology is applied across numerous fields of life.
The ethical and safety issues concerning the use of recombinant DNA techniques must be carefully analyzed and addressed before implementation.