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Pune University - Instrumentation and Molecular Biophysics, April 2010

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Total No. of Questions : 5] [Total No. of Pages : 2 P1009 [3725] - 201 M.Sc. MICROBIOLOGY MB - 601 : Instrumentation and Molecular Biophysics (2008 Pattern) Time : 3 Hours] [Max. Marks : 80 Instructions to the candidates: 1) All questions are compulsory. 2) All questions carry equal marks. 3) Draw neat labeled diagrams wherever necessary. 4) Figures to the right indicate full marks. 5) Use of logarithmic tables, electronic pocket calculator is allowed. 6) Assume suitable data if necessary. Q1) Attempt any two of the following : [16] a) Describe the construction and working of Gas Chromatography. How does the peak height get affected with change in split ratio? b) Give applications of Gel Filtration Chromatography. Does the shape of proteins affect the elution and why? c) Explain the principle of Fluorescence Spectroscopy. Describe use of Fluorescence Spectroscopy in binding studies and protein folding studies. Q2) Attempt any two of the following : [16] a) Explain the instrumentation required for X-ray Crystallography. What are planes in a crystal? Explain with diagram the Miller Indices of (100) in a simple cuboid. b) What is the principle of NMR Spectroscopy? Explain the terms chemical shift coupling constant and NOE. c) Give the basic principle of Mass spectrometry. Explain how a TOF instrument works. P.T.O. Q3) Attempt any two of the following : [16] a) Explain the concept of Ramchandran plot. How do various angles in polypeptide chain decide the structure of proteins? b) How are the prediction of secondary structures done by the ChouFasman method? State how it is different from GOR method. c) Give the principle of Tracer Technique. Give applications of Tracers in biology. Q4) Write short notes on any four of the following : a) Iso-electric focusing. b) X-ray Diffraction. c) Isopycnic Centrifugation. d) Beer Lambert Law and its limitations. e) Super Secondary structures of protein. Q5) Solve : a) i) b) [16] [16] Predict the order of elution when a mixture containing the following compounds is passed through a column containing a gel that excludes all proteins of molecular weight 200,000 and higher : cytochrome c (Mr = 13,000), tryptophan synthetase (Mr = 17,000), hexokinase (Mr = 96,000), ATP sulfurylase (Mr = 440,000), glucose oxidase (Mr = 154,000), and xanthine oxidase (Mr = 300,000). ii) What factors other that molecular weight will influence the elution volume of a protein from a Sephadex column? A solution containing 10-5M ATP has a transmission 0.702 (70.2%) at 260 nm 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. vvvv [3725]-201 2

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Additional Info : M.Sc. MICROBIOLOGY, MB - 601 : Instrumentation and Molecular Biophysics (2008 Pattern), Pune University
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