Genetic Engineering Applications and Processes

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Questions and Answers

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?

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?

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?

Industrial applications Signup and view all the answers

Which PCR technique is specifically designed for quantifying gene expression?

Quantitative real-time PCR (qPCR) Signup and view all the answers

What is the primary function of vectors in biotechnology?

To transfer and propagate foreign DNA fragments Signup and view all the answers

Which field is NOT traditionally associated with the goals of genetic engineering?

Cosmetic surgery advancements Signup and view all the answers

What is a key characteristic of expression vectors?

They ensure stable expression of the gene of interest. Signup and view all the answers

What is the significance of isolating a target gene in cloning processes?

To manipulate and observe gene expression variations Signup and view all the answers

Which of the following correctly describes pseudo-colonies in the context of cell competency?

They can represent artifacts or satellite colonies. Signup and view all the answers

What characterizes high copy number plasmids like pUC19?

They have a relaxed control mechanism. Signup and view all the answers

Which of the following factors can influence plasmid yield?

All of the above Signup and view all the answers

What is true about low copy number plasmids like pSC101?

They operate under stringent control. Signup and view all the answers

Which bacterial strain characteristic can affect plasmid stability?

Presence of specific endonucleases Signup and view all the answers

How does the insert affect plasmid replication?

Repetitive sequences can destabilize the plasmid. Signup and view all the answers

What is a key advantage of using E.coli as a prokaryotic host in genetic engineering?

It can grow well in low nutrition. Signup and view all the answers

Which organism is suggested to potentially replace E.coli as a host in genetic engineering?

Vibrio natriegens Signup and view all the answers

What method is commonly used to induce competency in E.coli for transformation?

Chemical method with CaCl2 and heat shock Signup and view all the answers

What characteristic of Vibrio natriegens is notable compared to E.coli?

It reproduces in less time than E.coli. Signup and view all the answers

What is the significance of a shuttle vector in genetic engineering?

It allows for expression in both prokaryotic and eukaryotic cells. Signup and view all the answers

Why are scientists skeptical about the research regarding Vibrio natriegens' potential to replace E.coli?

The study used a non-peer-reviewed platform for publication. Signup and view all the answers

Which of the following describes the structure of E.coli's genome?

It is fully sequenced and has 4.6 million base pairs. Signup and view all the answers

What is an example of a physical method to induce competency in E.coli?

High electric field electroporation Signup and view all the answers

What is the primary function of the Origin of replication (Ori) in a plasmid?

It ensures that the vector can self-replicate in the host cell. Signup and view all the answers

Which of the following components is NOT a basic part of an artificial plasmid?

RNA polymerase gene Signup and view all the answers

What role do colicin factors play in plasmids?

They serve as an anti-competitor agent. Signup and view all the answers

What is a cryptic plasmid?

A plasmid with no observable effect on the host cell. Signup and view all the answers

Which type of plasmid typically has a high copy number?

High copy number plasmid. Signup and view all the answers

Which of the following statements accurately describes plasmids?

Plasmids replicate independently of chromosomal DNA. Signup and view all the answers

Resistance transfer factors in plasmids are responsible for which function?

Transferring resistance traits to antibiotics. Signup and view all the answers

What is the significance of having high A-T and low G-C content in the Ori?

It may influence the replication speed. Signup and view all the answers

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.