Selected Questions in Molecular Biology

  1.  Avery, McCleod, and McCarty provided evidence that the substance responsible for converting a nonpathogenic form of pneumococci into a pathogenic form is DNA, not proteins or other cellular constituents. Explain with the help of diagrams.

  2. How are oncogenes acquired? Give the possible mechanisms.

    ·  Introduction by viral infection is one possible route. RNA retroviruses can accidentally incorporate RNA originating from mRNA for a control gene. If this RNA leads to the production of an oncogenic product then a tumour will result.

    ·  The normal cellular equivalent is called a proto-oncogene. Examples of oncogenes include those derived from ras (v-ras) and from erb (v-erb).

    ·  Oncogenes can also be acquired via changes in the cellular proto-oncogene so that it becomes an oncogene directly. This can occur by mutation of chromosomal rearrangement.

    ·  Sometimes the normal proto-oncogene becomes overproduced by being under the control of an inappropriate promoter/enhancer and this leads to oncogenesis.

  3. If a gene that is inserted into a vector is to be expressed, what other components must be included?

  4. Can all human proteins be produced relatively easily using bacteria once we have isolated the human gene in cDNA form? Explain. 

  5. If you were attempting to produce a human protein using recombinant techniques in a bacterial system would you use genomic DNA? Why? 

  6. Translation is the mechanism of protein synthesis. Some newly synthesised proteins are directly secreted out through ER. How are these proteins secreted through the ER membrane? 

  7. How many high-energy phosphates does it require for the direct synthesis of a protein of 200 amino acids in length? Do not include the cost of making the components required, such as amino acids, nor the cost of making the protein synthetic apparatus.

  8. Would you expect tRNAs within a given organism to be the same size or can they vary? Why?

  9. Would you expect the insertion of a single base into a gene or changing a single base within a gene to have more effect? Why?

  10. Newly synthesised proteins during translation are prevented from folding until it reaches its destination. What mechanism prevents these newly synthesised proteins from folding and getting in to the functional 3D form. Explain.

  11. Mitochondrion is a eukaryotic organelle. But it has its own DNA and protein synthesising machinery. List the major differences between the protein synthesising machinery of mitochondrion and the nuclear genome.
  12. How does protein synthesis in eukaryotes differ from that of prokaryotes? Explain.

  13. How is fidelity of translation achieved? Explain the possible mechanism to incorporate the correct amino acids as per the codon. 

     

     

  14.  Explain the various steps in the molecular mechanism of peptide elongation during protein translation.

  15. Explain the various molecular events that leads to the initiation of translation in E.coli.

  16.  Ribosomes are considered as the platform of protein translation. Explain how the structure and  chemical composition of ribosomes are related to its function.

  17.  The transfer of the exact amino acids to the site of protein synthesis is the function of tRNA. The attachment of the amino acid top the respective tRNA is known as ‘charging of tRNA’ and the charged tRNA is known as amino acyl tRNA. Explain the molecular mechanisms that leads to the formation of aminoacyl tRNA. How the amino acids recognise its respective tRNA during the process of ‘charging’.

  18. The effect of certain point mutations can be nullified by the mechanism of wobble pairing. Explain.

  19. Explain the secondary structure of transfer RNA with a neatly drawn diagram.

  20. Draw the genetic code and give the names of amino acids, which has one and two codons.  Which are the stop codons?

  21. Explain what is meant by the term ‘polysome’ and how polysomes arise.
  22.  Compare protein synthesis in mitochondria and in prokaryotes.

  23. Describe the steps involved in the initiation of protein synthesis in bacteria.

  24. Outline the process of peptide chain termination during translation.
  25. Explain why protein synthesis is a target for many inhibitors and provide at least three examples of inhibitors and their site of action.

  26. Histone genes have the following characteristics: they do not have introns, are arranged in multiple tandem arrays each with one copy of the five histone genes and do not have poly A tails. What explanation can be suggested for these features?

  27. What position you can give for ribozymes in the evolution of DNA as the genetic material? Explain.

  28. There are some genes that do not code for a protein. Explain its importance and its functions.
  29. Write a short note on peptidyle transferase.
  30. Describe the mechanism of gene transcription in eukaryotes, showing how the production of the final mRNA differs from that which occurs in prokaryotes.

  31. What is the need for the transcription of genes that do not code for a protein? What are the RNA polymerases that are involved in their transcription?
  32. Discuss the possible reasons for the existence of split genes in eukaryotes and the way in which they are removed from the mRNA prior to its transport to the cytoplasm.
  33. Explain the various post transcriptional modification of the primary transcript, which leads to the formation of mature mRNA in eukaryotes.
  34. Describe the structure of eukaryotic genes upstream from the transcription start site.
  35. Describe the mechanism of formation of the basal initiation (transcription) complex in eukaryotes.
  36. Explain the mechanism of transcription termination in eukaryotes.
  37. What is meant by splicing? Alternative splicing can lead to the production of two (or more) proteins for the price of one gene. Explain.

  38. Give a detailed account on RNA polymerases of eukaryotes and its division of labour with respect to the types of genes to be transcribed.
  39.  Describe transcription processes in mitochondria, emphasizing how these differ from nuclear processes.

  40. What are the factors that control mRNA stability and control of gene expression.

  41. Give a detailed account of various types of eukaryotic promoters and its role in the transcriptional regulations.

  42. Explain what is meant by the term ‘ribozyme’ and its role in the production of mature mRNA in eukaryotic systems. Give at least one example.
  43. What are the arguments put forward for the view that the first living material was RNA based?

  44. Why are there multiple genes for rRNA but only single copy genes for ribosomal proteins?

  45. RNA polymerase has no proofreading activity but DNA polymerase does. Why the difference? Explain

  46. What are Zinc finger proteins? Explain its importance in the gene regulation.

  47. Describe the structure of RNA and explain how it differs from DNA.

  48. Define the terms transcription, translation, template strand and coding strand in the context of molecular biology.

  49. List some of the general properties of prokaryotic mRNA
  50. The following is the sequence of an mRNA. Give the sequence of the coding strand or sense strand of its gene. AUC GCC UAG AGC CCC UUC CCA AGG CGC CUG GGC UGA AAA AAA

  51. Explain the mechanism of attenuation.

  52. Draw a schematic diagram of the structure of the lac operon and explain the functions of the various components. Describe the role of the lac operon in the life of E. coli.
  53. Using the tryptophan operon (or a similar operon) as an example compare and contrast control of a repressible operon with that of an inducible operon.

  54. Explain mechanisms of termination of transcription in prokaryotes.

  55. Explain the various molecular mechanisms leading to the formation of initiation complex during the process of transcription in prokaryotes.
  56. Explain organization and mechanism of action of his Operon. How it is different from lac operon and trp operon.
  57. What is sigma factor? Provide examples of different forms of sigma factor and their biological roles.

  58. What are the various types of DNA binding proteins involved in the regulation gene expression in both eukaryotes and prokaryotes?

  59. What is meant by Shine–Delgarno sequence ? Where it is located? Explain its role in gene expression.

  60. What is the possible relevance of the existence of telomeres and the occurrence of telomerase to profound biological events such as cancer and ageing?

  61. Cells treated with UV light have a higher survival rate if they are exposed to light immediately afterwards? Explain.

  62. RNA synthesis does not involve proof reading as in the case DNA synthesis.Does the use of an RNA primer for DNA synthesis affect the accuracy of DNA replication in E. coli? Explain.

  63. In many strains of bacteria the doubling time is less than the time taken to synthesise a complete copy of the DNA. How might the bacteria cope with this situation? Does a similar situation arise in animals?

  64. Explain the molecular mechanism of homologous recombination E.coli.

  65. What are transposons or mobile genes? Explain its nature, different classes and its biological   significance.

  66. The genomes of some retroviruses have homology with that human DNA sequences. Explain the possible origin of retroviruses on the basis of transposons.

  67. Transposons are mutagens. Explain this statement with the help of evidences.

  68. What are LINEs, SINEs and MITEs? Explain.
  69. Explain how telomerase acts to overcome the problem associated with the replication of the ends of DNA.

  70. Explain the various DNA repair mechanisms which exist in eukaryotes.
  71. List differences between eukaryote and prokaryote DNA synthesis.

  72. Explain the meaning of ‘semiconservative replication’ in the context of DNA. Show how the Meselson and Stahl experiment provided evidence if support of this mechanism of replication.

  73. What is mutation? What are the various types of mutations and explain the molecular mechanisms. Explain frame shift mutation. 

  74. Show how looping of DNA permits 5' ® 3' synthesis of the lagging strand.

  75. What are the various enzymes and proteins that involved in the process of DNA replication in prokaryotes?

  76. What are topoisomerases?  What are the biological implications of topoisomerases?

  77.  Define the terms ‘relaxed’, ‘positively supercoiled’ and ‘negatively supercoiled’ and show how these are relevant to DNA replication.

  78.  Discuss the role of negative supercoiling in DNA function, including transcription.

  79.  Explain the polarity problem in DNA replication and how this is overcome.

  80.  Explain the mechanism of methyl-directed mismatch repair.

  81. Give a comparative account of various types of DNA polymerase in prokaryotes? Explain the role of each enzyme DNA replication and repair.  

  82. List differences between eukaryote and prokaryote DNA replication.

  83. Explain various DNA repair mechanisms, which operate in E. coli with examples.

  84. What are Okazaki fragments. Explain how it could solve the problem of polarity during DNA replication.

  85. Explain the Klenove fragment. What is its practical application in molecular biology experiments? Explain.

  86. Explain the terms ‘split genes’, ‘pseudo genes’ ‘overlapping genes’ and ‘cryptic genes ‘.

  87. Differentiate between overlapping gene and polycistronic gene. What is the advantage of polycistronic genes?

  88. What are the basic enzymic reaction catalysed by DNA polymerases during DNA replication.
  89. Describe the way in which initiation of DNA synthesis is controlled in E. coli.
  90. Define the terms ‘relaxed’, ‘positively supercoiled’ and ‘negatively supercoiled’ and show how these are relevant to DNA replication.

  91. What is a replication fork? Explain about the enzyme complex present at the replication fork in E.coli.

  92. Explain the mechanism of termination of DNA replication in bacteria.

  93. What is Xeroderma pigmentosum? How it caused.

  94. Explain the molecular mechanism of SOS response.

  95. What are the biological implications of topoisomerases?
  96. Explain the mechanism of methyl-directed mismatch repair.

  97. Describe the events occurring to remove RNA, proofread and ligate DNA.

  98. Histone genes have the following features: a) no introns; b) the genes are arranged in tandem arrays with one copy of each gene per group; c) the mRNAs have no poly A tail. What explanation can be offered for these properties?

  99. Give a comparative account on the eukaryotic DNA polymerases and bacterial polymerases, including its structural organization and function.

  100. Explain various mechanisms of gene regulation in eukaryotes.

  101. Compare the structure and organization of eukaryotic genes with that of bacteria.

  102. What are transcription factors (TF)? Explain how they are involved in the regulation of gene expression with respect to tissue, cells and age with suitable examples.

 

 

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