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Questions and Answers
What is the main difference between microinjection and biolistic transformation?
What is the primary purpose of phage introduction in genetic engineering?
Who developed methods for splitting DNA molecules at selected sites and attaching segments to the DNA of a virus or plasmid?
What is the fundamental goal of laboratory geneticists?
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Which technology forms the basis of recombinant DNA technology?
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What is the process of transfection in genetic engineering equivalent to?
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What is another term for Recombinant DNA as mentioned in the text?
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Which process involves combining the DNA of two different organisms?
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What is the purpose of a selectable marker in Recombinant DNA?
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Which enzyme is used to cut a piece of DNA during the process of making Recombinant DNA?
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What happens to a host cell without a vector when exposed to a certain antibiotic?
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Which organism is given as an example of a possible host cell in the text?
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Study Notes
Recombinant DNA Basics
- Recombinant DNA is a general term for combining a piece of one DNA with another strand of DNA, also referred to as "chimera".
- The most common recombinant process involves combining the DNA of two different organisms.
Recombinant DNA Methods
- There are three methods to make Recombinant DNA: Transformation, Phage Introduction, and Non-Bacterial Transformation.
Transformation Method
- Select a piece of DNA to be inserted into a vector.
- Cut the DNA with a restriction enzyme.
- Ligate the DNA insert into the vector with DNA Ligase.
- The inserted DNA contains a selectable marker, often an antibiotic marker, to identify recombinant molecules.
- The vector is inserted into a host cell, which is a process called transformation.
- E.Coli is an example of a possible host cell.
Non-Bacterial Transformation Method
- In microinjection, the DNA is injected directly into the nucleus of the cell being transformed.
- In Biolistic, the host cells are bombarded with high-velocity micro-projectiles, such as particles of gold or tungsten, coated with DNA.
- Non-bacterial transformation does not use bacteria such as E.Coli as the host.
Phage Introduction Method
- In vitro packaging of a vector is used to produce phage plaques containing recombinant DNA.
- Lambda or MI3 phages are used to produce phage plaques.
- Recombinants can be identified by differences in the recombinants and non-recombinants using various selection methods.
Historical Background
- Werner Arber, a Swiss microbiologist, discovered restriction enzymes in 1968.
- Hamilton O. Smith purified Type II restriction enzymes in 1969.
- Paul Berg developed methods for splitting DNA molecules at selected sites and attaching segments of the molecule to the DNA of a virus or plasmid.
- Stanley N. Cohen and Herbert W. Boyer were the first to insert recombined genes into bacterial cells in 1973.
Background Information
- Recombinant DNA technology is the joining of DNA molecules from two different species to produce new genetic combinations.
- The goal of laboratory geneticists is to isolate, characterize, and manipulate genes.
- Recombinant DNA technology is based on cloning and DNA sequencing.
- Cloning is undertaken to obtain the clone of one particular gene or DNA sequence of interest.
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Description
Learn about the fundamentals of Recombinant DNA, including its process and applications. This quiz covers topics like combining DNA strands from different organisms and methods for creating Recombinant DNA.
