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Pune University - Biophysics, Instrumentation & Bioinform, April 2010 (Old Course)

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Total No. of Questions : 5] [Total No. of Pages : 3 P1002 [3725] - 33 M.Sc. MICROBIOLOGY MB - 703 : Biophysics, Instrumentation and Bioinformatics (2005 Pattern) Time : 3 Hours] [Max. Marks : 80 Instructions to the candidates: 1) All questions are compulsory. 2) All questions carry equal marks. 3) Neat well labeled diagrams must be drawn wherever necessary. 4) Use of log tables and electronic pocket calculators is allowed. 5) Assume suitable data if necessary. Q1) Attempt any two of the following: [16] a) Explain the principle behind protein gel electrophoresis. Differentiate between native and SDS gel electrophoresis. b) Explain the principle behind gel filtration chromatography. What will be effect of column size on the separation of proteins? How can we determine the molecular weight of the protein using gel filtration chromatography? c) Explain the working and applications of gas liquid chromatography. Q2) Attempt any two of the following : [16] a) Explain isomerism with respect to the structures of amino acids. Which amino acid is an exception? Draw Fisher projection for an L-amino acid. b) Describe the construction and working of MALDI TOF. Explain the concept of time of flight. c) What is nuclear resonance spectroscopy (NMR)? Explain the terms Chemical Shift and Spin-spin coupling in NMR. P.T.O. Q3) Attempt any two of the following : [16] a) Explain neural networks method for protein secondary structure determination. b) Explain the dynamic programming method for pair wise sequence alignment. How will you align two sequences ATTGC and AGGC if the identical match is assigned a score of 1, mismatch a score 0 and gap penalty is - 1? c) What are scoring matrices? How do they help in finding similarity between proteins? Q4) Write short notes on any four of the following : a) Anomalous scattering in X-ray diffraction. b) Homology based protein structure determination. c) Dynamic programming for sequence alignment. d) Relative Centrifugal Force. e) [16] Pulse chase experiment. Q5) Solve : a) [16] A solution containing 10-5 M ATP has a transmission 70.2% at 260nm in a 1 cm cuvette. Calculate the i) Transmission of the solution in a 3cm cuvette. ii) Absorbance of the solution in the 1cm and 3cm cuvettes and iii) Absorbance and transmission of a 5 10-5M ATP solution in a 1cm cuvette. b) You have a mixture of proteins with the following properties : Protein 1 : MW 12,000, Protein 2 : MW 62,000, pI = 4 Protein 3 : MW 28,000, pI = 8 Protein 4 : MW 9,000, [3725]-33 pI = 10 pI = 5 2 Predict the order of elution of these proteins when a mixture of four is chromatographed in the following system : i) DEAE cellulose at pH 7, with a linear salt gradient elution. ii) CM cellulose at pH 7, with a linear salt gradient elution. iii) A gel exclusion column with a fraction range of 1,000 - 30,000 MW at pH 7. vvvv [3725]-33 3

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Additional Info : M.Sc. MICROBIOLOGY, MB - 703 : Biophysics, Instrumentation and Bioinformatics (2005 Pattern), Pune University
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