REVISION on Recomb. DNA (1).pdf

Revision on Recombinant DNA Technology Lec.1 What is recombinant DNA? o A) DNA from a single source o B) A molecule that combines DNA from two sources o C) DNA that is degraded o D) DNA that is synthesized in a laboratory o Answ...

Revision on Recombinant DNA Technology Lec.1 What is recombinant DNA? o A) DNA from a single source o B) A molecule that combines DNA from two sources o C) DNA that is degraded o D) DNA that is synthesized in a laboratory o Answer: B) A molecule that combines DNA from two sources Which major project is associated with the advancement of recombinant DNA technology? o A) Apollo Mission o B) Human Genome Project o C) Manhattan Project o D) CERN's Large Hadron Collider o Answer: B) Human Genome Project What is the primary benefit of placing the human gene for insulin in bacteria? o A) To create a new species o B) To produce insulin in large quantities for diabetics o C) To eliminate diabetes o D) To study bacterial genetics o Answer: B) To produce insulin in large quantities for diabetics What universal property of the genetic code makes genetically modified organisms possible? o A) Variability in genetic codes o B) Differences in species' genetic codes o C) The universal nature of the genetic code o D) The uniqueness of each organism's genetic code o Answer: C) The universal nature of the genetic code What is the first step in recombinant DNA technology? o A) Joining DNA fragments o B) Isolating the gene of interest o C) Cloning the DNA o D) Screening the recombinant DNA o Answer: B) Isolating the gene of interest What is the term used for the introduction of recombinant DNA into host cells? o A) Transformation o B) Transcription o C) Translation o D) Transduction o Answer: A) Transformation What is the role of restriction endonucleases in recombinant DNA technology? o A) To replicate DNA o B) To cut DNA into fragments 1  o C) To ligate DNA fragments o D) To isolate DNA o Answer: B) To cut DNA into fragments In the context of recombinant DNA technology, what does 'ligation' refer to? o A) The replication of DNA o B) The cutting of DNA o C) The joining of DNA fragments o D) The transcription of DNA o Answer: C) The joining of DNA fragments What is the purpose of cloning the DNA of interest into a vector? o A) To degrade the DNA o B) To produce multiple copies of the DNA o C) To prevent the DNA from replicating o D) To create mutations in the DNA o Answer: B) To produce multiple copies of the DNA What is the final step in the recombinant DNA technology process as described in the document? o A) Cutting the DNA o B) Inserting DNA into bacterial cells o C) Screening the recombinant DNA o D) Recovering and analyzing the cloned DNA o Answer: D) Recovering and analyzing the cloned DNA Lec. 4 , manipulating DNA 1. What is the primary purpose of gene cloning experiments? o A) To destroy unwanted DNA o B) To construct recombinant DNA molecules o C) To modify RNA molecules o D) To create single-stranded DNA 2. What are the two examples of DNA manipulative techniques mentioned? o A) Shortening and lengthening DNA o B) Cutting and joining DNA o C) Copying and modifying DNA o D) Degrading and repairing DNA 3. Which enzyme is involved in degrading DNA molecules by breaking phosphodiester bonds? o A) Ligases o B) Polymerases o C) Nucleases o D) Modifying enzymes 4. What is the function of ligase enzymes in DNA manipulation? o A) To degrade nucleic acid molecules o B) To make copies of molecules o C) To join nucleic acid molecules together o D) To add or remove chemical groups 2 5. Which enzyme is commonly used in the polymerase chain reaction (PCR)? o A) DNA polymerase I o B) Taq DNA polymerase o C) Reverse transcriptase o D) Polynucleotide kinase 6. What type of enzyme is used to remove phosphate groups from the 5′ terminus of a DNA molecule? o A) Alkaline phosphatase o B) Polynucleotide kinase o C) Terminal deoxynucleotidyl transferase o D) Reverse transcriptase 7. Which type of nuclease removes nucleotides one at a time from the end of a DNA molecule? o A) Endonucleases o B) Exonucleases o C) Ligases o D) Polymerases 8. Which endonuclease enzyme is purified from the fungus Aspergillus oryzae? o A) Bal31 o B) Exonuclease III o C) S1 endonuclease o D) Deoxyribonuclease I (DNase I) 9. What is the role of DNA ligase during DNA replication? o A) To synthesize new DNA strands o B) To repair single-stranded breaks in DNA o C) To degrade RNA molecules o D) To add chemical groups to DNA 10. Which enzyme is used to synthesize a DNA strand complementary to an RNA template? o A) Taq DNA polymerase o B) DNA polymerase I o C) Reverse transcriptase o D) Terminal deoxynucleotidyl transferase 11. How many different restriction endonucleases have been isolated? o A) Over 1000 o B) Over 1500 o C) Over 2000 o D) Over 2500 12. Which type of restriction endonucleases are most important in gene cloning? o A) Type I o B) Type II o C) Type III o D) Type IV 13. What does the enzyme DNase I target in a DNA molecule? o A) Specific sequences o B) Only single strands 3  o C) Any internal phosphodiester bond o D) Only double-stranded regions 14. Which enzyme attaches to a short single-stranded region in a mainly double- stranded DNA molecule? o A) DNA polymerase I o B) Taq DNA polymerase o C) Reverse transcriptase o D) Alkaline phosphatase 15. What are sticky ends in DNA manipulation? o A) Blunt ends of DNA molecules o B) Ends of DNA that can form base pairs with complementary sequences o C) Ends of DNA that cannot be ligated o D) Single-stranded regions within a double-stranded molecule 16. What is the function of the Klenow fragment? o A) To degrade RNA o B) To add phosphate groups o C) To synthesize new DNA strands without degrading existing ones o D) To cleave specific DNA sequences 17. Which enzyme is thermostable and used in PCR? o A) DNA polymerase I o B) Taq DNA polymerase o C) Reverse transcriptase o D) Polynucleotide kinase 18. What is the source organism for Taq DNA polymerase? o A) Escherichia coli o B) Thermus aquaticus o C) Calf thymus tissue o D) Alteromonas espejiana 19. Which enzyme can add deoxyribonucleotides onto the 3′ terminus of a DNA molecule? o A) DNA polymerase I o B) Polynucleotide kinase o C) Terminal deoxynucleotidyl transferase o D) Alkaline phosphatase 20. What are linkers in DNA manipulation? o A) Enzymes that cut DNA o B) Short pieces of double-stranded DNA with a known sequence o C) Enzymes that ligate DNA fragments o D) Proteins that bind to DNA 21. What problem can arise when using linkers in DNA manipulation? o A) They can form dimers o B) They require high temperatures to function o C) They can cleave the target DNA molecule o D) They cannot be synthesized in large amounts 22. What is the difference between linkers and adaptors? o A) Linkers have sticky ends while adaptors do not 4  o B) Adaptors have one sticky end pre-formed while linkers are blunt-ended o C) Linkers are synthetic while adaptors are natural o D) Adaptors are used for ligation while linkers are used for cleavage 23. How is homopolymer tailing achieved in DNA manipulation? o A) By adding linkers to DNA fragments o B) By synthesizing short double-stranded DNA sequences o C) By using terminal deoxynucleotidyl transferase to add nucleotides to the 3′-OH termini o D) By cutting DNA with restriction endonucleases 24. What is the result of a partial digestion in restriction mapping? o A) Complete degradation of DNA o B) Cleavage at every possible restriction site o C) A complex pattern of bands in an electrophoresis gel o D) No cleavage of the DNA molecule 25. What is the main purpose of restriction mapping? o A) To sequence DNA molecules o B) To determine the size of DNA fragments o C) To illustrate the positions of recognition sequences in DNA o D) To synthesize new DNA strands 26. What type of gel is typically used for separating large DNA molecules in electrophoresis? o A) Polyacrylamide gel o B) Agarose gel o C) Denaturing gel o D) Non-denaturing gel 27. How does orthogonal field alternation gel electrophoresis (OFAGE) differ from standard gel electrophoresis? o A) It uses a continuous electric field o B) It uses a pulsed field alternating between two pairs of electrodes o C) It requires a higher voltage o D) It separates DNA molecules based on charge rather than size 28. What is the significance of the migration rate of DNA molecules during electrophoresis? o A) It is directly proportional to the size of the molecules o B) It is inversely proportional to the size of the molecules o C) It remains constant for all molecule sizes o D) It increases with the complexity of the DNA structure 29. Which enzyme removes nucleotides from both strands of a double-stranded DNA molecule? o A) S1 endonuclease o B) DNase I o C) Bal31 exonuclease o D) Exonuclease III 30. Which enzyme is involved in the replication of viruses and uses RNA as a template? o A) DNA polymerase I o B) Taq DNA polymerase 5  o C) Reverse transcriptase o D) Terminal deoxynucleotidyl transferase 31. How can the sizes of DNA fragments in a gel be estimated most accurately? o A) By using a standard molecular weight marker o B) By comparing with previously known fragment sizes o C) By using a mathematical relationship linking migration rate to molecular mass o D) By visual inspection 32. What modification do bacterial DNA undergo to protect it from restriction endonucleases? o A) Phosphorylation o B) Methylation o C) Dephosphorylation o D) Glycosylation 33. What is the primary role of restriction endonucleases in bacteria? o A) To synthesize new DNA strands o B) To degrade foreign DNA o C) To replicate bacterial DNA o D) To repair damaged DNA 34. What does the process of ligation involve in DNA manipulation? o A) Breaking phosphodiester bonds o B) Joining two DNA fragments o C) Removing nucleotides o D) Adding chemical groups to DNA 35. Which enzyme is responsible for adding phosphate groups to the 5′ terminus of a DNA molecule? o A) Alkaline phosphatase o B) Polynucleotide kinase o C) Terminal deoxynucleotidyl transferase o D) Reverse transcriptase 36. What characteristic is required for an enzyme to be useful in a PCR reaction? o A) It must be able to join DNA fragments o B) It must be thermostable o C) It must degrade RNA o D) It must have exonuclease activity 37. Which of the following is a technique for separating DNA molecules based on size using an electric field? o A) Ligation o B) Restriction mapping o C) Gel electrophoresis o D) Homopolymer tailing 38. What is the purpose of using a restriction enzyme in DNA manipulation? o A) To synthesize RNA from DNA o B) To cut DNA at specific sequences o C) To join DNA fragments together o D) To add nucleotides to DNA 39. What is the result when DNA is partially digested with a restriction enzyme? 6  o A) All possible cleavage sites are cut o B) Only a subset of the possible cleavage sites are cut o C) No cleavage occurs o D) The DNA is fully degraded 40. What enzyme would you use to repair single-stranded breaks in DNA? o A) Taq DNA polymerase o B) DNA ligase o C) Reverse transcriptase o D) DNase I Model Answer 1. What is the primary purpose of gene cloning experiments? o B) To construct recombinant DNA molecules 2. What are the two examples of DNA manipulative techniques mentioned? o B) Cutting and joining DNA 3. Which enzyme is involved in degrading DNA molecules by breaking phosphodiester bonds? o C) Nucleases 4. What is the function of ligase enzymes in DNA manipulation? o C) To join nucleic acid molecules together 5. Which enzyme is commonly used in the polymerase chain reaction (PCR)? o B) Taq DNA polymerase 6. What type of enzyme is used to remove phosphate groups from the 5′ terminus of a DNA molecule? o A) Alkaline phosphatase 7. Which type of nuclease removes nucleotides one at a time from the end of a DNA molecule? o B) Exonucleases 8. Which endonuclease enzyme is purified from the fungus Aspergillus oryzae? o C) S1 endonuclease 9. What is the role of DNA ligase during DNA replication? o B) To repair single-stranded breaks in DNA 10. Which enzyme is used to synthesize a DNA strand complementary to an RNA template? o C) Reverse transcriptase 11. How many different restriction endonucleases have been isolated? o C) Over 2000 12. Which type of restriction endonucleases are most important in gene cloning? o B) Type II 13. What does the enzyme DNase I target in a DNA molecule? o C) Any internal phosphodiester bond 14. Which enzyme attaches to a short single-stranded region in a mainly double- stranded DNA molecule? o A) DNA polymerase I 15. What are sticky ends in DNA manipulation? 7  o B) Ends of DNA that can form base pairs with complementary sequences 16. What is the function of the Klenow fragment? o C) To synthesize new DNA strands without degrading existing ones 17. Which enzyme is thermostable and used in PCR? o B) Taq DNA polymerase 18. What is the source organism for Taq DNA polymerase? o B) Thermus aquaticus 19. Which enzyme can add deoxyribonucleotides onto the 3′ terminus of a DNA molecule? o C) Terminal deoxynucleotidyl transferase 20. What are linkers in DNA manipulation? o B) Short pieces of double-stranded DNA with a known sequence 21. What problem can arise when using linkers in DNA manipulation? o A) They can form dimers 22. What is the difference between linkers and adaptors? o B) Adaptors have one sticky end pre-formed while linkers are blunt-ended 23. How is homopolymer tailing achieved in DNA manipulation? o C) By using terminal deoxynucleotidyl transferase to add nucleotides to the 3′-OH termini 24. What is the result of a partial digestion in restriction mapping? o C) A complex pattern of bands in an electrophoresis gel 25. What is the main purpose of restriction mapping? o C) To illustrate the positions of recognition sequences in DNA 26. What type of gel is typically used for separating large DNA molecules in electrophoresis? o B) Agarose gel 27. How does orthogonal field alternation gel electrophoresis (OFAGE) differ from standard gel electrophoresis? o B) It uses a pulsed field alternating between two pairs of electrodes 28. What is the significance of the migration rate of DNA molecules during electrophoresis? o B) It is inversely proportional to the size of the molecules 29. Which enzyme removes nucleotides from both strands of a double-stranded DNA molecule? o C) Bal31 exonuclease 30. Which enzyme is involved in the replication of viruses and uses RNA as a template? o C) Reverse transcriptase 31. How can the sizes of DNA fragments in a gel be estimated most accurately? o A) By using a standard molecular weight marker 32. What modification do bacterial DNA undergo to protect it from restriction endonucleases? o B) Methylation 33. What is the primary role of restriction endonucleases in bacteria? o B) To degrade foreign DNA 34. What does the process of ligation involve in DNA manipulation? o B) Joining two DNA fragments 8 35. Which enzyme is responsible for adding phosphate groups to the 5′ terminus of a DNA molecule? o B) Polynucleotide kinase 36. What characteristic is required for an enzyme to be useful in a PCR reaction? o B) It must be thermostable 37. Which of the following is a technique for separating DNA molecules based on size using an electric field? o C) Gel electrophoresis 38. What is the purpose of using a restriction enzyme in DNA manipulation? o B) To cut DNA at specific sequences 39. What is the result when DNA is partially digested with a restriction enzyme? o B) Only a subset of the possible cleavage sites are cut 40. What enzyme would you use to repair single-stranded breaks in DNA? o B) DNA ligase 9 MCQs on Protein Purification for Recombinant DNA Question 1: Which of the following is the first step in the purification of recombinant proteins? A) Affinity chromatography B) Gel electrophoresis C) Cell lysis D) Dialysis Answer: C) Cell lysis Question 2: What is the main purpose of using affinity chromatography in protein purification? A) To separate proteins based on their size B) To separate proteins based on their charge C) To specifically bind and isolate the target protein D) To precipitate proteins out of solution Answer: C) To specifically bind and isolate the target protein Question 3: Which tag is commonly used for purifying recombinant proteins using affinity chromatography? A) His-tag B) GST-tag C) FLAG-tag D) All of the above Answer: D) All of the above Question 4: In ion-exchange chromatography, proteins are separated based on their: A) Size B) Charge C) Hydrophobicity D) Solubility Answer: B) Charge 10 Question 5: What is the role of SDS in SDS-PAGE? A) To maintain the native structure of proteins B) To impart a uniform negative charge to the proteins C) To cleave proteins at specific sites D) To act as a buffer Answer: B) To impart a uniform negative charge to the proteins Question 6: Which method can be used to remove salts from a protein solution after ion-exchange chromatography? A) Centrifugation B) Dialysis C) Gel filtration chromatography D) Both B and C Answer: D) Both B and C Question 7: During protein purification, what is typically monitored to track the presence of the target protein? A) pH of the solution B) Protein concentration using a spectrophotometer C) Enzymatic activity or specific binding assay D) Conductivity of the solution Answer: C) Enzymatic activity or specific binding assay Question 8: Which type of chromatography would be most appropriate for separating proteins based on their hydrophobicity? A) Affinity chromatography B) Ion-exchange chromatography C) Hydrophobic interaction chromatography D) Size-exclusion chromatography Answer: C) Hydrophobic interaction chromatography 11 Question 9: In the purification of His-tagged proteins, which molecule is commonly used in the elution buffer? A) Imidazole B) Glutathione C) EDTA D) NaCl Answer: A) Imidazole Question 10: Which technique is often used as a final step in protein purification to achieve high purity and homogeneity? A) Ammonium sulfate precipitation B) Affinity chromatography C) Gel filtration chromatography D) Differential centrifugation Answer: C) Gel filtration chromatography Protein Gel Electrophoresis What is the main purpose of gel electrophoresis? A) To mix proteins in a solution B) To separate molecules based on size and charge C) To synthesize new proteins D) To visualize DNA sequences Answer: B) To separate molecules based on size and charge SDS-PAGE is primarily used to separate proteins based on what property? A) Their charge B) Their size C) Their shape D) Their color Answer: B) Their size What role does SDS (sodium dodecyl sulfate) play in SDS-PAGE? 12  A) It adds color to the proteins B) It maintains the pH of the solution C) It denatures proteins and gives them a uniform charge D) It catalyzes the polymerization of acrylamide Answer: C) It denatures proteins and gives them a uniform charge What is the purpose of the stacking gel in SDS-PAGE? A) To polymerize acrylamide B) To stain the proteins C) To concentrate the proteins into tight bands D) To denature the proteins Answer: C) To concentrate the proteins into tight bands Which component in the sample preparation buffer helps to break disulfide bonds in proteins? A) Tris buffer B) Glycerol C) SDS D) β-mercaptoethanol Answer: D) β-mercaptoethanol What is the effect of using a continuous buffer system in gel electrophoresis? A) It provides higher resolution separation B) It results in fuzzy and unresolved protein bands C) It speeds up the polymerization process D) It increases the size of the gel pores Answer: B) It results in fuzzy and unresolved protein bands What is the most common dye used for staining proteins after electrophoresis? A) Ethidium Bromide B) Crystal Violet C) Coomassie Blue D) Safranin Answer: C) Coomassie Blue What is the role of APS (Ammonium Persulfate) in the preparation of polyacrylamide gels? 13  A) To stain the proteins B) To act as a reducing agent C) To catalyze the polymerization of acrylamide D) To maintain the pH of the solution Answer: C) To catalyze the polymerization of acrylamide How does the concentration of acrylamide in the resolving gel affect protein separation? A) Higher concentration results in larger pores B) Lower concentration results in smaller pores C) Higher concentration results in smaller pores D) It does not affect pore size Answer: C) Higher concentration results in smaller pores What is the primary driving force behind protein folding as mentioned in the presentation? A) Covalent bonding B) Hydrogen bonding C) Hydrophobic interactions D) Electrostatic interactions Answer: C) Hydrophobic interactions 14  Applications of recombinant DNA Technology What is recombinant DNA technology primarily used for in biopharmaceuticals? A) Synthesizing inorganic compounds B) Cloning animals C) Producing proteins and hormones D) Mapping human genomes Answer: C) Producing proteins and hormones Which of the following is a key benefit of using recombinant DNA technology in vaccine development? A) Producing vaccines in a shorter time B) Eliminating the need for clinical trials C) Reducing the cost of raw materials D) Increasing the shelf life of vaccines Answer: A) Producing vaccines in a shorter time Which bacterium is commonly used as a host for producing recombinant proteins? A) Staphylococcus aureus B) Bacillus subtilis C) Escherichia coli D) Mycobacterium tuberculosis Answer: C) Escherichia coli Human insulin produced using recombinant DNA technology is known as: A) Humulin B) Actrapid C) Novolin D) Insuman Answer: A) Humulin Which of the following is an example of a recombinant vaccine? A) Polio vaccine B) Hepatitis B vaccine C) Tuberculosis vaccine D) Influenza vaccine 15 Answer: B) Hepatitis B vaccine What role does plasmid DNA play in recombinant DNA technology? A) It acts as a vector to transfer genes B) It codes for ribosomal RNA C) It degrades unwanted DNA D) It synthesizes proteins directly Answer: A) It acts as a vector to transfer genes Which enzyme is used to cut DNA at specific sequences in recombinant DNA technology? A) DNA polymerase B) Ligase C) Restriction endonuclease D) Reverse transcriptase Answer: C) Restriction endonuclease The introduction of recombinant DNA into host cells is known as: A) Transcription B) Transformation C) Translation D) Transduction Answer: B) Transformation Recombinant human growth hormone (rhGH) is used to treat: A) Diabetes B) Growth hormone deficiency C) Anemia D) Hepatitis B Answer: B) Growth hormone deficiency Which of the following is NOT a step in creating recombinant DNA? A) Isolating the gene of interest B) Inserting the gene into a host genome randomly C) Ligating the gene into a vector D) Amplifying the gene using PCR Answer: B) Inserting the gene into a host genome randomly 16  What is the main advantage of recombinant vaccines over traditional vaccines? A) They are cheaper to produce B) They are always more effective C) They are safer and cause fewer side effects D) They require no storage Answer: C) They are safer and cause fewer side effects In biopharmaceutical production, what is a commonly used mammalian cell line? A) E. coli B) Yeast cells C) HeLa cells D) CHO cells (Chinese Hamster Ovary cells) Answer: D) CHO cells (Chinese Hamster Ovary cells) Which of the following describes a biopharmaceutical? A) A small molecule drug B) A product derived from living organisms C) A synthetic chemical compound D) An inorganic mineral supplement Answer: B) A product derived from living organisms Recombinant DNA technology can be used to produce monoclonal antibodies for: A) Diagnostic purposes B) Treating infections C) Cancer therapy D) All of the above Answer: D) All of the above Which method is commonly used to introduce recombinant DNA into plant cells? A) Microinjection B) Agrobacterium-mediated transformation C) Electroporation D) Liposome fusion Answer: B) Agrobacterium-mediated transformation 17  What is the purpose of using a selectable marker gene in recombinant DNA experiments? A) To identify cells that have taken up the recombinant DNA B) To degrade non-recombinant DNA C) To enhance the replication of plasmids D) To visualize DNA fragments Answer: A) To identify cells that have taken up the recombinant DNA Which of the following biopharmaceuticals is produced using recombinant DNA technology? A) Aspirin B) Insulin C) Paracetamol D) Ibuprofen Answer: B) Insulin Gene therapy, an application of recombinant DNA technology, aims to: A) Replace defective genes with functional ones B) Repair damaged tissues C) Enhance athletic performance D) Alter physical appearance Answer: A) Replace defective genes with functional ones What is a potential advantage of DNA vaccines over traditional vaccines? A) They can be stored at room temperature B) They do not require adjuvants C) They can induce both humoral and cellular immunity D) They are effective against all diseases Answer: C) They can induce both humoral and cellular immunity Which type of recombinant protein is commonly used as a treatment for multiple sclerosis? A) Recombinant insulin B) Interferon-beta C) Human growth hormone D) Erythropoietin 18 Answer: B) Interferon-beta 19