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Molecular Biotechnology

Subject : engineering
Instructions:
  • Answer 50 questions in 15 minutes.
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  • Match each statement with the correct term.
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This is a study tool. The 3 wrong answers for each question are randomly chosen from answers to other questions. So, you might find at times the answers obvious, but you will see it re-enforces your understanding as you take the test each time.
1. Comprised of >50 proteins associated with rRNA units. Site of protein synthesis and binds mRNA and finds protein synthesis initiation sites. It also binds aa- tRNA and catalyzes peptide bond formation.






2. Genes for products that are required at all times.






3. 1. Ethidium bromide staining 2. P32 - P33 radioactivity 3. Fluorescence 4. Agarose gel electrophoresis






4. 1. Capping: 5' phosphate capped by 7- methyl guanosine and is a 5'-5' linkage instead of 5'-3' This makes RNA more stable 2. Intron removal 3. Export to cytoplasm 4. Polyadenylated mRNA precursor






5. The small ribosomal subunit binds to 5'-G cap on processed mRNA (no RBS) - uses met instead of fmet for initiation; monocistronic translation






6. Operons transcribed as single mRNA and mRNA codes for more than one protein.






7. Release DNA - rewind DNA - release RNA; stop signals or rho mediated termination (hairpin is a palindromic GC- rich region followed by an AT- rich region; Rho is a termination factor that binds to nascent RNA) RNAP has sigma factor that recognizes pr






8. The process of increasing the expression of inducible genes






9. The process in which an exact copy of the double strand DNA is made. It is a templated process and occurs from 5' to 3' end. DNAP - RNA primer; semiconservative (each strand is a template for the replication of the complementary strand)






10. A cell that contains a nucleus and membrane bound organelles






11. Replication > DNA > Transcription > RNA > Translation > Protein






12. In the presence of glucose and lactose - bacteria grows first on glucose - then growth levels off - and starts growing on lactose. You have diauxie growth because (1) CAP helps recruit RNAP. in the presence of glucose - CAMP is low so it can't bind t






13. Important to suppress mutations at 3rd position and you don't need to have a lot of stop codons; cells can be more flexible






14. A reading frame without termination codon among 50 or more codons. Usually correspond to genes that encode proteins






15. Production of commercial products generated by the metabolic actions of microorganisms.






16. 1. Initiation: unwind DNA at the origin of replication (ori) - bidirectional replications; regulated as required for cell division 2. Elongation: requires RNA primer to replicate 3. Termination: signaled by Ter sequence






17. A haploid organism that is diploid for a small region of the chromosome (partial diploid)






18. Ribosome doesn't stop at trp codons and stem loop forms between 3 and 4. RNAP stops prematurely (attenuated)






19. A small catabolite molecule. Its level is determined by the level of glucose in the cell where glucose controls the rate of cAMP formation with ATP. When there is high glucose - there is low levels of cAMP. cAMP activator protein (CAP) has to bind cA






20. Polymerase binds to lac promoter weakly by itself and results in low levels of transcription even in the absence of lacI. The activator recruits the polymerase to the promoter region and increases its affinity for the promoter






21. Attenuation






22. Three sites recruit tRNA and forms peptidyl- tRNA bonds (E - exit; P - peptide; A - acceptor).






23. The first two bases of the codon always form strong Watson -Crick base- pairing. The first base in the anticodon determines the number of codons a tRNA can recognize. The first position in anticodon is often modified to inosine to facilitate wobble b






24. A segment of DNA molecule contains the information required for synthesis of a functional biological product






25. Search for site to start transcription - unwind DNA; -35 region and pribnow region (-10 region).






26. Start codon is usually ATG - first amino acid is n - formyl- methionine. It is assisted by initiation factors (IF) and requires ribosomal binding sites (RBS). It is a polycistronic protein translation (operon).






27. Determines amino acid selection. A noncognate amino acid charge incorrectly to the tRNA will be inserted into the protein. Introduce new amino acid by using tRNA for UAG.






28. Multiple effects from a single gene






29. 1. Nucleic acid hybridization: (a) bind single stranded DNA to a membrane support - (b) add single stranded labeled DNA (probe) under appropriate conditions - (c) wash the support to remove excess unbound labeled probe DNA - (d) detect the hybrid seq






30. When arabinose is present - it binds to araC and allosterically induces it to bind to araI instead araO2. If glucose is absent - then the presence of CAP bound to its site between araO1 and araI helps break the DNA loop and helps araC bind to araI






31. Operator site; araC bound at this site can simultaneously bind to the araI site to repress transcription from Pbad promoter






32. Codes for three enzymes needed to catalyze the metabolism of arabinose. The operon is regulated by araC gene product.






33. Functions: enzymes - regulation - structural - cellular functions Polymers of amino acids and connected by peptide bonds. Can fold into complex structures.






34. 4. Cells + organelle 3. Supermolecular complexes 2. Macromolecules 1. Monomeric units






35. In E. coli - DNAP III can unwind DNA (helicase) and replicate both strands of DNA. It also has proofreading activity and corrects mistakes 3' to 5' exonuclease






36. Reverse Transcriptase






37. When half DNA strand has been denatured. Determined by GC content (triple bond)






38. Operator site - araC binds to this site and represses its own transcription from the PC promoter. In the presence of arabinose - araC bound at this site helps to activate expression of Pbad promoter






39. LacY: Transports lactose into the cell LacZ: B- galactosidase LacA: transacetylase LacI: lacI+ cells fully inducible - lacI- were already induced and not responsive to IPTG X- gal: analog of lactose that turns blue when cleaved by lacZ product and o






40. Structural and functional units of life. All organisms are made of cells - all cells are derived from preexisting cells - the purpose of a microorganism is to make another microorganisms as quickly as possible; alter metabolism of microorganism to ma






41. When arabinose is absent - there is no need to express the structural genes. AraC does this by binding simultaneously to araI and araO2 - making a looped DNA. This blocks access to Pbad promoter. AraC is an autoregulator of its own expression and the






42. Nonsense mutation in gene that results in truncated protein can be lethal. Sometimes a second mutation arises that counteracts the effects of the mutation. Amber stop codon (UAG/TAG/etc) and amber suppressor tRNA (CUA/etc) can restore protein size an






43. Gene products decrease in concentration under particular molecular circumstances






44. Gene products increase in concentration under particular molecular circumstances






45. 1. Synthesis of commercial products by recombinant organisms 2. Biopolymers 3. Bioremediation 4. Biomass utilization






46. 1. LacI- makes an internal inducer -- NO. Found that lacI- doesn't dominate over lacI+ and is not always constitutive. 2. LacI- is a repressor protein -- YES. LacI+ dominates over lacI- because when both are together - lac operon is inducible. LacI m






47. The ribosome translating the leader peptide arrives at the two tryptophan codons and has to wait for tryptophan. During this time - RNAP continues to transcribe. Stem loop between 2 and 3.






48. Inducer site; araC bound at this site can simultaneously bind to the araO2 site to repress transcription from the Pbad promoter. In the presence of arabinose - araC bound at this site helps to activate expression of Pbad promoter.






49. Chromosome (contains host genetic information) - plasmids (prokaryotes; small - self- replicating DNA; supercoil) - free nucleotides






50. The repressor dimer (aporepressor) can't bind to the repressor. Transcription from the promoter is not stopped. When tryptophan is bound to the repressor dimer - the repressor changes configuration so that it can bind to the operator and transcriptio