Podcast
Questions and Answers
What is the primary goal of recombinant DNA technology?
What is the primary goal of recombinant DNA technology?
- To revert an organism to its original genetic state
- To create genetically identical organisms
- To decrease the genetic diversity of a population
- To produce a desired product by modifying an organism’s DNA (correct)
Which of the following is a method used in gene manipulation?
Which of the following is a method used in gene manipulation?
- Inhibiting gene expression permanently
- Random mutation without guidance
- Isolation and cloning of DNA (correct)
- Selective breeding of non-altered organisms
In genetic engineering applications, what is the significance of vectors?
In genetic engineering applications, what is the significance of vectors?
- They are used to destabilize gene functions
- They cause mutations in the host DNA
- They transport genetic material into host cells (correct)
- They solely replicate without any modifications
Which aspect of genetic engineering is focused on increasing or decreasing specific gene expression?
Which aspect of genetic engineering is focused on increasing or decreasing specific gene expression?
Which PCR technique is specifically designed for quantifying gene expression?
Which PCR technique is specifically designed for quantifying gene expression?
What is the primary function of vectors in biotechnology?
What is the primary function of vectors in biotechnology?
Which field is NOT traditionally associated with the goals of genetic engineering?
Which field is NOT traditionally associated with the goals of genetic engineering?
What is a key characteristic of expression vectors?
What is a key characteristic of expression vectors?
What is the significance of isolating a target gene in cloning processes?
What is the significance of isolating a target gene in cloning processes?
Which of the following correctly describes pseudo-colonies in the context of cell competency?
Which of the following correctly describes pseudo-colonies in the context of cell competency?
What characterizes high copy number plasmids like pUC19?
What characterizes high copy number plasmids like pUC19?
Which of the following factors can influence plasmid yield?
Which of the following factors can influence plasmid yield?
What is true about low copy number plasmids like pSC101?
What is true about low copy number plasmids like pSC101?
Which bacterial strain characteristic can affect plasmid stability?
Which bacterial strain characteristic can affect plasmid stability?
How does the insert affect plasmid replication?
How does the insert affect plasmid replication?
What is a key advantage of using E.coli as a prokaryotic host in genetic engineering?
What is a key advantage of using E.coli as a prokaryotic host in genetic engineering?
Which organism is suggested to potentially replace E.coli as a host in genetic engineering?
Which organism is suggested to potentially replace E.coli as a host in genetic engineering?
What method is commonly used to induce competency in E.coli for transformation?
What method is commonly used to induce competency in E.coli for transformation?
What characteristic of Vibrio natriegens is notable compared to E.coli?
What characteristic of Vibrio natriegens is notable compared to E.coli?
What is the significance of a shuttle vector in genetic engineering?
What is the significance of a shuttle vector in genetic engineering?
Why are scientists skeptical about the research regarding Vibrio natriegens' potential to replace E.coli?
Why are scientists skeptical about the research regarding Vibrio natriegens' potential to replace E.coli?
Which of the following describes the structure of E.coli's genome?
Which of the following describes the structure of E.coli's genome?
What is an example of a physical method to induce competency in E.coli?
What is an example of a physical method to induce competency in E.coli?
What is the primary function of the Origin of replication (Ori) in a plasmid?
What is the primary function of the Origin of replication (Ori) in a plasmid?
Which of the following components is NOT a basic part of an artificial plasmid?
Which of the following components is NOT a basic part of an artificial plasmid?
What role do colicin factors play in plasmids?
What role do colicin factors play in plasmids?
What is a cryptic plasmid?
What is a cryptic plasmid?
Which type of plasmid typically has a high copy number?
Which type of plasmid typically has a high copy number?
Which of the following statements accurately describes plasmids?
Which of the following statements accurately describes plasmids?
Resistance transfer factors in plasmids are responsible for which function?
Resistance transfer factors in plasmids are responsible for which function?
What is the significance of having high A-T and low G-C content in the Ori?
What is the significance of having high A-T and low G-C content in the Ori?
Study Notes
Methodology of Genetic Engineering
- Genetic Engineering Applications:
- Agriculture: Production of desired products, manipulation of genes for increased/decreased expression.
- Industry: Modification of an organism's DNA.
- Medicine: Improvement of treatment methods.
- Genetic Engineering Process:
- Isolate and clone a gene
- Recombinant DNA technology
- Gene delivery to target tissue
- Prokaryotic Host: E. coli
- Advantages: Easy to cultivate, rapid replication, fully sequenced genome, extensive knowledge, many strains available.
- Disadvantages: Potential for toxicity.
- Alternative Host: Vibrio natriegens
- Replicates faster than E. coli
- Gene Manipulation:
- Vectors: Tools used to transfer foreign DNA into host cells.
- Shuttle Vectors: Vectors capable of replication in multiple hosts.
- Host Cell Competency
- Methods: Chemical (CaCl2 + heat shock), Physical (electroporation).
- Troubleshooting Competent Cells:
- Pseudo-colonies: Satellite or artifact colonies that are usually non-transformants.
- Selecting the Target Gene:
- Methods: Genomic DNA library, cDNA library, PCR, RT-PCR.
- Vectors: Carriers for Propagation and Expression:
- Cloning Vectors: Ensure propagation of target genes in the host cells.
- Expression Vectors: Ensure stable expression of genes in the host cells.
- Plasmids: Circular DNA Molecules in Bacteria:
- Self-replicating, can carry hundreds of genes.
- Types: High-copy number (smaller size), Low-copy number (larger size).
- Functions: Sex Factors, Colicin Factors, Resistance Transfer Factor.
- Artificial Plasmids (Vectors):
- Key Components: Origin of Replication (Ori), Multiple Cloning Site (MCS), Selectable Marker, Promoter, Sequence Elements of Translation.
- Vector Elements:
- Origin of Replication (Ori): Enables self-replication in the host cell.
- Multiple Cloning Site (MCS): Region for inserting target genes.
- Selectable Marker: Gene that confers resistance to an antibiotic, allowing selection of transformed cells.
- Promoter: Controls gene expression by determining transcription initiation.
- Sequence Elements of Translation: Enable proper translation of the target gene.
- Origin of Replication (Ori) Types:
- High Copy Number: Replicates independently of host chromosome, results in high vector copy number per cell.
- Low Copy Number: Replicates synchronously with host chromosome, resulting in lower vector copy number per cell.
- Plasmid Copy Number Control:
- Insert size: Toxic proteins or repetitive sequences can impact copy number.
- Bacterial Strain: Endonucleases can degrade plasmids.
- Growth Conditions: Temperature, nutrients, etc., can influence plasmid copy number.
- Culture Inoculum: Fresh inoculum is essential.
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Description
This quiz focuses on the methodology of genetic engineering, covering its applications in agriculture, industry, and medicine. It discusses the genetic engineering process, including gene isolation, recombinant DNA technology, and the use of different hosts like E. coli and Vibrio natriegens. Test your knowledge on gene manipulation and host cell competency techniques.
