p.9
Applications of DNA Technology in Medicine
What is the purpose of RFLP analysis?
To observe whether an individual carries a mutant gene for a hereditary disease.
p.6
Detection of DNA Sequences
How do hybridization probes assist in DNA analysis?
They allow the production of sufficient amounts of recombinant DNA for further analysis.
p.1
Gene Expression Regulation at Translation Level
Can one miRNA regulate multiple polypeptides?
Yes, one miRNA can block translation of several different polypeptides.
p.3
Clinical Examples of Deregulation of Gene Expression
What is the association between homocysteine and cardiovascular disease?
Increased concentration of homocysteine in blood is associated with a higher risk of atherosclerosis and thrombosis, leading to cardiovascular disease.
p.3
Clinical Examples of Deregulation of Gene Expression
What are some important oxidative modifications of proteins during aging?
Formation of dityrosines, oxidation of tryptophan, and reaction of free amino groups of lysine residues with aldehydes.
p.11
Production of Therapeutic Proteins
What is cloned DNA used to produce?
Hormones (e.g., insulin), therapeutic proteins (e.g., antibodies), and vaccines.
What is gene manipulation?
The use of biotechnology to produce different bioproducts by introducing new characteristics into recipient organisms.
p.1
Gene Expression Regulation at Translation Level
What is the significance of the 5‘-UTR and 3‘-UTR in translation regulation?
They contain sequences that serve as binding sites for regulatory proteins, influencing translation efficiency.
p.1
Gene Expression Regulation at Translation Level
How does binding to the 3‘-UTR affect translation?
It increases the efficiency of translation mainly due to increased stability of mRNA.
p.7
Detection of DNA Sequences
What is the role of labeled antibodies in Western blotting?
To visualize the protein of interest.
p.10
Gene Therapy Techniques
What types of viruses can be used in gene therapy?
Retrovirus, adenovirus, lentivirus, and herpes simplex virus.
p.4
Methods in DNA Technology
What is the first step in preparing a DNA library?
Cutting the investigated DNA into fragments using restriction endonuclease.
p.10
Clinical Examples of Deregulation of Gene Expression
How is sickle cell anemia diagnosed?
Using restriction fragment length polymorphism (RFLP).
p.9
Applications of DNA Technology in Medicine
How is probe-target hybridization detected in DNA arrays?
By detecting and quantifying fluorophore-labeled targets.
p.5
Recombinant DNA Technology
What is the purpose of recombinant DNA technology?
To introduce new, favorable characteristics into organisms, such as resistance to insect attack.
p.10
Detection of DNA Sequences
What is DNA fingerprinting primarily used for in forensic medicine?
Identification of criminals, murder victims, or for paternity tests.
p.8
Methods in DNA Technology
What are the two methods developed for DNA sequencing?
Maxam and Gilbert method, and Sanger’s method.
p.1
Gene Expression Regulation at Translation Level
How can the synthesis rate of a particular protein change?
It can change quickly due to alterations in metabolism and physiological conditions, influenced by regulatory protein binding.
p.8
Detection of DNA Sequences
What is in situ hybridization used for?
Detection of DNA or RNA sequences directly in investigated tissue.
p.8
Detection of DNA Sequences
What is the purpose of the labeled hybridization probe in in situ hybridization?
To target the sequence of interest.
p.2
Post-Translational Modifications of Proteins
What is dephosphorylation?
The removal of a phosphate group from proteins, catalyzed by phosphatases.
p.7
Detection of DNA Sequences
What is the purpose of hybridization probes?
To identify DNA library clones containing the sequence of interest.
p.6
Methods in DNA Technology
What happens during the DNA synthesis step of PCR?
The mixture is heated to 72°C for elongation of DNA from primers using thermostable DNA polymerase.
p.11
Gene Therapy Techniques
What is the primary use of recombinant DNA technology in therapy?
Preparation of peptides, proteins, antisense oligonucleotides, RNA interference, and gene therapy.
p.1
Gene Expression Regulation at Translation Level
What is the role of miRNA in translation regulation?
Binding of miRNA to target mRNA leads to quick elimination by ribonucleases and blocks translation elongation.
p.11
Gene Therapy Techniques
What is the role of miRNA and siRNA in RNA interference?
They direct enzyme complexes to degrade mRNA molecules, decreasing their activity and preventing translation.
p.9
Applications of DNA Technology in Medicine
In what fields is in situ hybridization used?
Prenatal diagnostics and oncology.
p.1
Gene Expression Regulation at Translation Level
What role do eIF-4E and eIF-4G play in translation initiation?
They bind to the CAP and internal ribosomal entry sites (IRES), which is essential for the initiation of translation.
p.7
Detection of DNA Sequences
How does Northern blotting differ from Southern blotting?
Northern blotting is used for the detection of RNA, while Southern blotting is for DNA.
p.7
Detection of DNA Sequences
What information does DNA sequencing provide?
The sequence of nucleotides in DNA.
p.11
Recombinant DNA Technology
What must an expression vector contain?
Replication origin and promoter sequences.
p.4
Recombinant DNA Technology
What is a clone in the context of recombinant DNA?
A population of host cells containing identical recombinant DNA.
p.4
DNA Libraries and Gene Manipulation
What does the term 'DNA library' refer to?
A collection of DNA fragments from one organism that have been cloned into vectors for identification and isolation.
p.4
DNA Libraries and Gene Manipulation
What is a genomic DNA library?
A collection of cloned DNA fragments from the whole genome of a cell.
p.8
Detection of DNA Sequences
What is the first step in in situ hybridization?
Fixing the tissue to avoid chemical changes.
p.4
Methods in DNA Technology
What methods can be used to identify the sequence of a gene of interest?
Hybridization probes or polymerase chain reaction (PCR).
p.3
Clinical Examples of Deregulation of Gene Expression
What is non-enzymatic glycation of proteins?
The reaction of plasma proteins with glucose, responsible for chronic complications of diabetes.
p.1
Gene Expression Regulation at Translation Level
How can posttranslational modifications affect eIF-4E and eIF-4G?
They can change the level of binding properties of these initiation factors, thus altering the rate of translation initiation.
p.1
Gene Expression Regulation at Translation Level
What effect does binding to the 5‘-UTR have on translation?
It is associated with inhibition of translation.
p.11
Gene Therapy Techniques
What is the purpose of antisense oligonucleotides?
To inhibit the synthesis of a target protein by reducing the level of specific mRNA.
p.8
Methods in DNA Technology
What is the basis of the Maxam and Gilbert method?
Chemical cleavage of DNA.
p.4
DNA Libraries and Gene Manipulation
What are the three kinds of DNA libraries?
Genomic DNA library, Chromosome DNA library, and cDNA libraries.
p.10
Gene Therapy Techniques
What are some risks associated with gene therapy?
Introduction of foreign objects, potential viral complications, and chance of inducing mutations.
p.2
Post-Translational Modifications of Proteins
What is the significance of ATP in the ubiquitin-proteasome pathway?
The attachment of ubiquitin to target proteins is ATP-dependent.
p.1
Gene Expression Regulation at Translation Level
What is the first mechanism of eukaryotic translation regulation?
Phosphorylation of eIF-2α by different protein kinases, leading to inhibition of translation initiation.
p.7
Detection of DNA Sequences
What are the main steps involved in Southern blotting?
Cutting genomic DNA, separating fragments by gel electrophoresis, transferring to a membrane, and hybridization with a probe.
p.9
Applications of DNA Technology in Medicine
What is Southern blotting used for?
To detect mutations and DNA insertions or deletions.
p.10
Gene Therapy Techniques
What methods are used for prenatal diagnosis?
PCR or in situ hybridization.
p.11
Gene Therapy Techniques
What is RNA interference (RNAi)?
A method of disease treatment that suppresses gene expression using double-stranded RNA.
p.4
DNA Libraries and Gene Manipulation
What is a cDNA library?
A collection of cloned DNA sequences that are transcribed into mRNA in a particular cell or tissue.
p.10
Clinical Examples of Deregulation of Gene Expression
What causes severe immuno-deficiency syndrome?
Mutation of the gene coding for adenine deaminase.
p.2
Post-Translational Modifications of Proteins
What are some other chemical modifications of polypeptide chains?
Methylation, acetylation, and hydroxylation.
p.7
Detection of DNA Sequences
What is Southern blotting used for?
Detection of DNA sequences.
p.6
Applications of DNA Technology in Medicine
What are some applications of PCR in human medicine?
Detection of mutations, molecular classification of tumors, identification of viruses or bacteria, microbiological screening of food, and DNA amplification in forensic medicine.
p.3
Clinical Examples of Deregulation of Gene Expression
What are prion diseases?
Neurodegenerative disorders associated with unusual post-translational modifications of prions.
p.4
Recombinant DNA Technology
What is recombinant DNA?
DNA composed of at least two parts from different organisms that can be replaced in a host organism.
p.5
Methods in DNA Technology
What is the role of DNA ligase in DNA technology?
To attach the DNA vector to the foreign DNA.
p.5
DNA Libraries and Gene Manipulation
What is DNA cloning?
A technique that makes many identical copies of a piece of DNA, such as a gene.
p.2
Post-Translational Modifications of Proteins
Give examples of proteins that undergo proteolytic activation.
Pepsinogen → pepsin, fibrinogen → fibrin, prothrombin → thrombin.
p.2
Post-Translational Modifications of Proteins
What is ubiquitin?
A small peptide that labels proteins for degradation by the proteasome complex.
p.2
Post-Translational Modifications of Proteins
What is phosphorylation?
The addition of a phosphate group to proteins, often catalyzed by kinases.
p.9
Applications of DNA Technology in Medicine
What is one practical application of recombinant DNA technology in medicine?
Diagnostics of human diseases.
What is the first step in gene manipulation?
Isolation of DNA fragments from the donor.
p.7
Detection of DNA Sequences
What is the purpose of Western blotting?
To detect proteins using labeled antibodies.
p.7
Detection of DNA Sequences
What can DNA sequencing be used for?
Localization of regulatory and gene sequences, comparison of homologous genes, and identification of mutations.
p.10
Gene Therapy Techniques
What is gene therapy?
The treatment of inherited diseases by delivering a normal copy of a defective gene.
p.2
Post-Translational Modifications of Proteins
What is proteolytic activation of proteins?
The conversion of inactive polypeptides into active proteins through proteolytic cutting.
p.2
Post-Translational Modifications of Proteins
What is the role of the 26S proteasome complex?
It removes aged and damaged proteins from the cell through proteolytic degradation.
p.4
Methods in DNA Technology
How are recombinant molecules introduced into a host bacterium?
Through a process called transformation.
p.6
Methods in DNA Technology
What is the first step in the PCR process?
Thermal denaturation, where primers are heated to 95°C to denature dsDNA into single strands.
What is a vector genome?
A DNA molecule that represents a small segment of DNA containing all signals necessary for replication.
p.6
Detection of DNA Sequences
What are probes in the context of DNA analysis?
Single stranded sequences of DNA or RNA that hybridize with complementary sequences of investigated DNA or RNA.
p.3
Clinical Examples of Deregulation of Gene Expression
What happens to normal prions in prion diseases?
Normal prions with dominant α-helix structure can misfold into aberrant prions with β-pleated sheets, leading to neurodegeneration.
p.7
Detection of DNA Sequences
What is the first step in Southern blotting?
Cutting genomic DNA by restriction endonuclease into discrete fragments.
p.11
Gene Therapy Techniques
How do antisense oligonucleotides work?
They bind to complementary coding regions on mRNA, forming RNA-DNA heteroduplexes that are cleaved by RNAse-H.
p.8
Methods in DNA Technology
What is the role of ddNTPs in Sanger’s method?
They prevent the addition of further dNTPs, terminating DNA chain synthesis.
p.5
Methods in DNA Technology
What are the three steps involved in one cycle of PCR?
Denaturation, annealing, and extension.
p.2
Post-Translational Modifications of Proteins
How does inadequate proteasome function relate to neurodegenerative diseases?
It can lead to the accumulation of aberrant proteins, contributing to diseases like Alzheimer’s and Parkinson’s.
p.9
Applications of DNA Technology in Medicine
What is a DNA array?
A high throughput method used to determine the expression level of many genes or mutations simultaneously.
p.9
Applications of DNA Technology in Medicine
What does RFLP stand for?
Restriction Fragment Length Polymorphism.
p.10
Detection of DNA Sequences
What is the basis of DNA fingerprinting?
The presence of variable number of tandem repeats (VNTR).
p.5
DNA Libraries and Gene Manipulation
How is a plasmid used in DNA cloning?
The target gene is inserted into a plasmid, which is then introduced into bacteria via transformation.
p.8
Detection of DNA Sequences
What is immunocytochemistry used for?
Detection and visualization of proteins or other antigens in cells using antibodies.
p.3
Clinical Examples of Deregulation of Gene Expression
What is glycated hemoglobin used for?
Long-term control of glycemia (blood glucose concentration).
p.3
Clinical Examples of Deregulation of Gene Expression
How does aging affect protein modifications?
Aging is associated with the slow accumulation of oxidized proteins, mainly in the extracellular matrix.
p.5
Methods in DNA Technology
What are restriction endonucleases?
DNA cutting enzymes used to cut fragments for the production of recombinant DNA.
p.6
DNA Libraries and Gene Manipulation
What was a significant use of hybridization probes?
For identification of DNA library clones containing the sequence of interest.
p.2
Post-Translational Modifications of Proteins
What are post-translational modifications?
Covalent modifications that significantly impact the function of modified proteins.
p.9
Applications of DNA Technology in Medicine
What does PCR stand for and what is its use?
Polymerase Chain Reaction; used for detecting DNA insertions, deletions, and gene amplification.
p.10
Clinical Examples of Deregulation of Gene Expression
What hereditary disease can be cured with ex vivo gene therapy?
Severe immuno-deficiency syndrome.
p.8
Detection of DNA Sequences
How do antibodies function in immunocytochemistry?
They specifically recognize the target of interest and are linked to a reporter.
p.3
Clinical Examples of Deregulation of Gene Expression
What is the consequence of prion misfolding?
It leads to the formation of protein aggregates that induce neuronal death and consequent neurodegeneration.
p.10
Gene Therapy Techniques
What is required for the normal gene to be effective in gene therapy?
It must be inserted into a specific vector for incorporation into the genomic DNA.
p.5
Methods in DNA Technology
What is the polymerase chain reaction (PCR)?
A method based on repetitive DNA synthesis using thermostable DNA polymerase and specific primers.
p.2
Post-Translational Modifications of Proteins
What is the role of protein kinase A?
It is activated by cAMP and plays a role in signal transduction.
p.8
Methods in DNA Technology
What is the key feature of Sanger’s method?
It uses labeled di-deoxy-nucleotides (ddNTPs) in addition to normal deoxynucleotides.
p.11
Gene Therapy Techniques
What are the two types of small RNA molecules involved in RNA interference?
MicroRNA (miRNA) and small interfering RNA (siRNA).
p.9
Applications of DNA Technology in Medicine
What is in situ hybridization used for?
To detect DNA or RNA sequences directly in tissue.
p.10
Clinical Examples of Deregulation of Gene Expression
What type of genetic inheritance is sickle cell anemia?
Autosomal recessive disease.
