Microbiology and Genetic Engineering Overview

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Questions and Answers

What is the purpose of using adaptors in molecular cloning?

To increase the size of plasmids

To promote the transformation of bacterial cells

To join different types of molecule ends (correct)

To enhance the stability of DNA molecules

What occurs during the brief heat shock in the transformation of E.coli?

The plasmids replicate within the cell

The DNA is completely degraded

The cell membrane becomes permeable to DNA (correct)

The E.coli undergoes cell division

What is the role of selectable markers in the transformation process?

To inhibit cell division in untransformed cells

To determine vector size

To facilitate gene expression

To identify successfully transformed cells (correct)

What happens when target DNA is inserted into the Tetr gene of a vector like pBR322?

Resistance to tetracycline is lost Signup and view all the answers

Which method is used to introduce vector DNA into host cells?

Transfection Signup and view all the answers

During transformation, which of the following is NOT a characteristic of untransformed host cells?

They can survive on antibiotic-containing media Signup and view all the answers

In the context of insertional selection, what does the presence of the ampr gene indicate?

The host cell can survive in ampicillin Signup and view all the answers

Which of the following best describes transfection?

Infection of bacteria by lambda phage particles containing recombinant DNA Signup and view all the answers

Which restriction enzyme is NOT present at the promoter of the tetracycline gene?

Eco RV Signup and view all the answers

What is the expected outcome when purified DNA is cut with Bam H1 and inserted into a phage vector?

Formation of circular DNA by pairing with the COS site Signup and view all the answers

Which type of vector contains two restriction sites flanking a replacement region?

λ replacement vectors Signup and view all the answers

What is the main advantage of using phage vectors over plasmid vectors for cloning?

Phages can handle larger DNA fragments than plasmids Signup and view all the answers

Which component is NOT a part of phage λ?

Replication aggressive factors Signup and view all the answers

In the lytic cycle, how does multiplication of the virus take place?

By cell lysis and infection of nearby cells Signup and view all the answers

What type of phage vector is represented by Lambda gt 10?

λ insertion vector Signup and view all the answers

What is a distinctive feature of a prophage?

It exists only in a lysogenic cycle. Signup and view all the answers

What characterizes monoclonal antibodies?

They are specific to a single antigenic determinant. Signup and view all the answers

What is the primary purpose of using monoclonal antibodies in laboratory tests?

To detect and quantify specific antibodies. Signup and view all the answers

Which of the following steps is NOT part of the hybridoma technology process?

Injecting multiple antigens for broad antibody production. Signup and view all the answers

In genetic engineering, what role do cloning vectors play?

They facilitate the replication of foreign DNA in host cells. Signup and view all the answers

Which enzyme is associated with hypoxanthine guanine phosphoribosyl transferase (HGPRT) activity?

It recycles components for RNA and DNA synthesis. Signup and view all the answers

What is the outcome of fusing B-lymphocytes with myeloma cells?

The formation of hybrid cells that can proliferate indefinitely. Signup and view all the answers

Which of the following uses of monoclonal antibodies does NOT pertain to diagnostics?

Production of vaccines. Signup and view all the answers

What is the term used for the process of making recombinant DNA?

Gene cloning. Signup and view all the answers

What is the primary function of the λ part in cosmids?

To aid in the packaging of DNA into the viral coat Signup and view all the answers

How are YACs different from typical plasmid vectors?

YACs contain elements required for chromosomal behavior Signup and view all the answers

Which feature of cosmids helps in the selection of cells containing the vector?

Markers for drug resistance Signup and view all the answers

What allows phage vectors to identify bacterial cells effectively?

Each phage particle forms a plaque in bacterial lawns Signup and view all the answers

Which component is NOT typically part of a YAC?

Bacteriophage packaging signal Signup and view all the answers

What determines the stability of linear YAC in yeast cells?

Inclusion of telomeres Signup and view all the answers

How does a virus insert genetic material into a host cell?

By infecting the host cell directly Signup and view all the answers

What is the maximum amount of DNA a cosmid can accommodate?

45kb Signup and view all the answers

What is a primary characteristic of continuous cell lines?

They can divide indefinitely and have no anchorage dependence. Signup and view all the answers

Which of the following processes is primarily used to produce cell suspensions for primary cell cultures?

Mechanical disaggregation of tissues. Signup and view all the answers

What is the role of DNA and RNA probes in microbiology?

To diagnose diseases such as kala azar and malaria. Signup and view all the answers

What type of cell culture is characterized by multiple generations and indefinite growth?

Continuous cell lines. Signup and view all the answers

In genetic engineering, what is one application of producing transgenic animals?

To study gene function and expression. Signup and view all the answers

What is the primary purpose of using artificial vaccines?

To prevent infectious diseases. Signup and view all the answers

Which method is commonly used in the detoxification of industrial effluents?

Cell culture technology. Signup and view all the answers

What is one of the features of finite cell lines?

They undergo a limited number of cell generations. Signup and view all the answers

Study Notes

DNA and RNA Probes

Utilized for diagnosing diseases like kala azar, sleeping sickness, and malaria.

Artificial Vaccines

Developed to stimulate immune response against specific pathogens.

Gene Therapy

A method for treating genetic diseases by correcting defective genes.

DNA Fingerprinting

A technique used for personal identification and forensic analysis.

Scope and Importance in Microbiology

Biochemical production including alcohol and antibiotics.
Protein and enzyme remodeling to enhance functionality.
Control of plant diseases and pest management (viruses, fungi, bacteria).
Industrial effluent and sewage treatment for detoxification.

Genetic Engineering

Involves manipulating genetic material to produce transgenic animals.
Enables rapid clonal multiplication of crops for agricultural improvement.
Focuses on developing virus-resistant and disease-resistant plant varieties.

Fundamentals of Animal Cell Culture and Hybridoma Technology

Cell culture allows for controlled cell growth in nutrient media.
Primary cell culture derived from directly isolated tissues.
Disaggregation techniques include enzymatic digestion and physical disruption.
Secondary culture involves subculturing from primary cultures.

Cell Lines

Defined as permanently established cell cultures with indefinite division potential.
Finite cell lines have limited division capability and show contact inhibition.
Continuous cell lines (transformed/cancerous) grow indefinitely, lacking density limitations.

Monoclonal Antibodies (mAb)

Produced from a single clone of B-cell, specific to a singular antigen.
Used for detecting antibodies, purifying substances, and diagnosing diseases.
Essential in immunohistochemistry and serological tests.

Hybridoma Technology Steps

Immunization of a rabbit leads to antibody production.
Fusion of B-lymphocytes with myeloma cells to produce hybrid cells.
Selected hybridoma cells are cultured for large-scale mAb production.

Gene Cloning and DNA Sequencing

Involves isolation and combination of DNA fragments for replication.
Requires cloning vectors (plasmid or phage) for DNA insertion and host transfer.

Phages

Bacterial-infecting viruses following lytic or lysogenic cycles.
Examples include λ phage and M13 with specific integration capabilities.

Vectors in Genetic Engineering

Phage vectors more efficient for large DNA cloning (20-23 kb).
Plasmid vectors accommodate less than 10 kb and require special introduction methods.
Cosmids combine features of plasmids and phage, accommodating large DNA inserts (up to 45 kb).

Yeast Artificial Chromosome (YAC)

Linear DNA segments that mimic yeast chromosomes, capable of cloning significant DNA (up to 1 Mb).
Contains necessary elements for replication, consists of left and right arms with telomeres.

Transformation and Transfection Methods

Transformation involves adding new DNA to bacterial cells, using calcium chloride and heat shock.
Transfection includes phage particles packaging recombinant DNA for infection into bacteria.

Detection of Recombinant Molecules

Selection methods utilize antibiotic resistance as a marker to distinguish transformed hosts.
Insertional selection inactivates certain antibiotic resistance genes to ensure accurate identification of recombinants.

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Description

Explore the essential concepts in microbiology and genetic engineering. This quiz covers DNA/RNA probes, artificial vaccines, gene therapy, and the importance of genetic manipulation in disease treatment and crop improvement. Test your knowledge on these vital topics that impact health and agriculture.