Biotechnology - Rayalaseema University

RAYALASEEMA UNIVERSITY:: KURNOOL (A.P.)M.Phil., / Ph.D., Written Examination SyllabusSub: BIOTECHNOLOGYPaper –I Research Methodology1. Cell and Tissue Culture Techniques:Isolation of Cells / tissues – Cell disruption techniques – Principles and application ofFrench press – Sonicator – Homogenizer – Precautions taken while disruption of cells –Cell fractionation – Isolation of periplasm – Cytoplasm and membranous tissue.Techniques involved in isolation of cell organelles – Nucleus, Mitochondria, Chloroplast– Density – gradient centrifugation. Sucrose and CsCI₂ density gradient centrifugation.2. Isolation Techniques:Nucleic acids – Isolation of genomic DNA from plane – Animal and microbial cells.Isolation of proteins from plant, animal and microbes. Separation of Methods - Cellfiltration – Ion exchange Chromatography – affinity Chromatography – HPLC –Principles and applications of HPLC, GLC – Participation Chromatography – PartitionCoefficient – Counter current distribution – Application of partition Chromatography inseparation and characterization of Biomolecules. Electrophoretic techniques forseparation and detection of DNA, RNA and proteins.3. Gene Manipulation Techniques:Isolation of genes from Prokaryotes and Eukaryotes – Chemical synthesis of DNA – invitro synthesis of gene – Restriction endonucleases – classification – Type II RE –Nomenclature – Enzymes that produce Protruding ends – 3 protruding ends – and theirsignificance in gene Vectors – Properties – Significance of Plasmid Vectors pBR322,pUC18/19, Bluecript constructed usuv for lacZ, tac, tric and T₇ promoters – Structure andfunctions Expression Vector – PET and PRSET Vectors – Expression of fusion proteins –Production of Proteins with Histag – Proteins – Significance. Ti-plasmid derived vectors– binary vectors – Significance in Molecular Cloning, BAC, YAC – Generation ofGenomic Libraries – in vitro chromosomal walking, Cloning in Animal Cells – Vectorsderived from SV40 – Creation of transgenic animals – Methods and Significance.4. Cloning Strategies:Cloning of DNA fragments with blunt and cohesive ended DNA Linker DNA –Homopolymer tailing. cDNA and genomic libraries. Transformation – Transduction –Transfection – Particle barbardment. Selection of Clones – Genetic – Immunochemical –1

Molecular Hybridization Methods. Protein Engineering – Principles of ProteinEngineering – site directed mutagenesis – Significance of M13 derive vectors –Engineering proteins fro thermo stability – resistance of high salt solution and detergents.DNA – Sequencing – Chemical enzymatic methods – Identification of regularityelements – reporter genes – use in identification of regulatory elements – Mapping oftranscriptional start site. PCR Techniques. Principles and applications.5. Radio Isotopes:Nature and properties of radioisotopes – Management of radioactivity – Safety aspects –Radio labeled precursor molecules – Preparation of probes – Neck translation, randomprime labeling – Purification of probes and measurement of specific activity. OpticalMethods – Spectroscopy – Microscopy – IR, ORD, CD, X-Ray diffraction techniquesand their signigicance in characterization of Biomolecules.2

RAYALASEEMA UNIVERSITY:: KURNOOL (A.P.)M.Phil., / Ph.D., Written ExaminationSub:BIOTECHNOLOGYPATTERN OF MODEL QUESTION PAPERTime : 3 hrs. Max. Marks 100Answer any FIVE questionsAll questions carry equal marks(Ten questions will be given selecting one from each chapter)123456789103