COURSE CO~TENTSBIOTECHNOLOGY

B.Se., r,il t& iit +e ~ ~E.~ur~OR. H.S.GOl:R U~I\,ERSITV, SAGAR 470003 M.P.

SCHEME OF EXAMINATiON

Class Papers I Practicals , MaximumMarks

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')()l. Biochemist}', Biochemical techniques &Biophysics

H. Microbial Technique & Rccornbincnt DNATechnique

Practical,· Project

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.t ~~~fft 50Methods in Molecular Biology, eel!

Biology & Animal Ceil Culture-Immunology & Immunological i kthods 50

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Biotechnology

"•..."".. ~ Ui1 !\j,dl1<;. Computers, EnvironmentalBi(~(i1C:lllSln and Cultural Methods

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COURSE CONTENTS IN BIOTECHNOLOGY

B.Sc.~ :r ~-e..,"~,,,l--tA.BIOCHEl\USTRY, BIOCHEltUCAL TECHNIQUES & BIO PHYSICS

lJNIT-I

Nature ofbiologicaJ material

(j ncral properties of'organic and inorganic compounds OJ

Sililabilityof organic compounds lor gCllcratiolllyfsll11cllm:, storage otenergy (U1J inlorm.uum 02f lydrophilic and hydrophobic groups In hiologicallllOlcclllcs UI('/;tssi fication ofbiomoleculcs based on their role ill hit iproccsses

vlolcculcs involved in ~encration 11fmcciJanicai stabi lity-pcptiJoglyeans,polysaccharid,« and IllcJllhral1~ lipids

Molecules imlllh'J in int<1l111atiollstorage and rctricval= thc nucleic acids

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iii Molecules executillg mediator and GJtaJ)'tic iunctions-the proteins1\. 111C signal rnolccutes.

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Perspectives ofhil)!ogical m<1cwll1olccu!cs: [he repeating units innucleic <Kids and proteins. Helicity, bending. looping. pleats. sail bridges cleo

;Illd their Jctcmlinants. lhe basis for irHCnl10kcllbr interaction e.g. CI1/YIllC-

substrate and ,lrltigl:.'l1-alltib()dy rCl'llgnili()ll,

; 'aturc ()fhiocilemic;d n:;Il'fio!l'i llndcrl:,ilh.! hiu,')~'llIhL'~is:lI1d dcgrad:lliol1.

Role prell/\. n!~'sill slIch tL';IClitlI]S. Pmtcin arid uon-protcin 1..'11/)111~S. Kinetics()r(,Il7~ me ellal) :il'll rl..'auit IllS,

III vitro acti\ it)" otpuri flL'dL'1l/\lTWS .md thc ir arplicatlt)Jl<; II1l1Jdtl~tr).

Various L1Sl'S\)(01/) I1h.',>- cl1/\ nics ill li,\,d pr\)L,:~:-'1Jl~, medicine, diat:l1osti-cs.md rWdlll'tiol1 I)I'IIL'\\ t·nll1pl111nJ::..

f·.1I/)11ICS as research tools - ELlS; methods, fl10dilicalinn ofblll/ogicaJcompouml-, \\ itb the help orcn/;' meso

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lnergclics nL,Ii\ ill,:! hu\l; :"lolll:Ccsot ht,.';Jl/illllt~ to 1I..-1/1I1C1:11111L'.

I lc.u di ..;~ip;lliull.lIld ~\lfN.'t\ Jtil 'I..

/,llJlhvl1-lkl..i" LJ\\_ Spl..·l.·lr(lpilllll'llh:ln. ,1110L"'ic)rillldrv

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Str~\1l.'sit:sollight n:cL'\)[I\JIl ill microbes, pL1I11S aJld '1I111l1;tj:.,

COITcclion ofvision faults[kclricll propcn i(~ () fhlological c<.\lllpal1llH.'nlSCiCI1l'r;lli\1I1 and nX'(-p11,!I1 lI!c;U111C vibrauons. l lc.mue didesI111ra-and intcrmolccul.u intcr.icuons in hi. dl)gieal ~\ <,ICI11S,S p:l1l;d ,lI1d l' h;lr)-',(

(pmp;ll ihi lit y :IS ILL1111in:1I11ofsuch interact Ions,

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tx IT-I\Physical methods appliL'd to lind out molecular structures

_ X-ray en sl.illographv and Nlvl R(jCl1cr~" spectroscopy -l\' - vis. lluo!i_:,>(\:ncc, :\\\\l11i( ,\hSOI1)\I()ll. IR ~\lld

Raman spectra.

Li\; IT-\'PI1\'sicalllh.:lh\,t! ...(li'IIl\;\~ill''': inl:1L'lhioioC'il..-a! structures.' .•... .•.... •..\ltr:hl)lllH.l, Oplil';t\ t't!ICIS. :\-1;..\)'. (',\ I scan, E(C.I":L(i ;lJJd :\\J1R ill1;lsin<>

B.Sc. i P€!f3efi' - II Se..W\f.-S+.e..~MICROBIAL TECHNIQUE & RECOMBINANT DNA TECHNIQUE

UNIT-IDevelopmcnl of microscopy (Optical. Tl·,~1.md SE\ 1} ()1Pastcu r"Sex Ix:ri Il1Cllts di ..;pnn i 11.:!,SpOllld!1":OUS gClleral i011 () 1ProL\l~\)tic (lnd ClI\.-;Il"\ olil' microhiul cells 02;\:tlllrc o fthc m 11.'rOhidi cell sllrb •...c. ( ir.un pl lsi I i \C and Gram-neg •.rti \'C ha •...tcria.

K intis of lbgclb, Sl"'"l \ lh::-; 0,2The various rCmll~()(1l1Ilr(lllr~:lIli ...;nl:' -PPl {):..;.l'IH'~·i. bacilli and spirilla U\

( \Illccpl of microbial "'pl'ci..::, ;JIId :..;lr;lin-. ()1'\ lit nIiOIl;JIl"I~hsilil''''ion I If I' lici PI )r~allisllb II I"\1'1'-11( ,,'lll'l il' hom. )~l'l 'L'i: y II I l'll I)1<1I populauon

1'11l11;1I1l'\'lIS alld il1dllCl·d variation arising in microhi.rl population\ I('IIC transfer in micu« irt:dll Isms

\ Illhi, )si~ ;!Old ;1111ihiosis among 11licrobial p\)PU \;II iullS\ 11(') nhcs in cxucmc environments - The t hcrmoph: k~ ;Illd :dk;l!t iphi ks1'.lllitl"Cll il' 1l11~'" II. )1',,:;1\II";\11,. Defence 111\.'( han i,.;md~:\ "':'111111. 10\ '1:.':llli :'11\";

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Microbial metabolism. Fermentation products. A survey of products frommicroorganisms. N !-fixing microbes in agriculture. 02

UNIT-IIIStrain improvement by enrichment, selection and recombinant DNA methods.Production ofhctcrologous protei ns of interest in microorganisms 04What is gene cloning and why do we need to clone a gene 02Tools and techniques, plasmids and other vehicles genomic DNA,handling of DNA. RNA, eDNA, RT enzymes and other reagents, techniques,laboratory req uirement 03Safety measures and regulations tor recomhinant'oNA work choice and selectionof the tools and the techniques. 02

UNIT-IVVehicles: Plasmids and bacteriophages, available phat!-cmids. cosmids, virusesPurification ofO!\iA from bacterial. plant and animal cellsManipulation of pur ill cd DNAIntroduction of DNA into liv ing cellsCloning vectors for EicoliCloning vectors for organisms other than F. coli yeast. lung], p lants-agrobact,plantvirus. animal viruses

UNIT-VApplication of cloning in gene analysis

How to obtain a clone ofa specific geneStudying gene location and structureStudying gene expression

Gene cloning and expression or foreign genes in research and biotechnologyProduction of protein from cloned genes

Gene cloning in medicinephannaccutical compoundsarti licial insul in gClk'

recombinant vaccinediagnostic rG1t!-Cllts.

PRACTICALSPart IQuantitative estimation of the following in biological samplesSugar in gi\ en solutionsSugar in biolo~:IL'<I1samplesExtraction .md separation ul' lipidsEst imat ion of protcms

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040203ll202

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I x3 hrsI x3 hrs2\3 hIS

2\,\ hI'S

Estimation of DNA/RNAAssay of enzyme activityKinetic studies on enzymesChromatographic methods for separation of macromolecules

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2x] hrs2x3 hrs5x3 Ius5x3 hrs

2x3 hrs2x3 hrs

2x3 hrs3x3 hrs2x3 hrs4x3 hrs3x3 hrs3x3 hrs2x3 hrs2x3 hrs

Part IIAseptic techniques:Cleaning of glasswares 2x3 hrsPreparation of media, cotton plugging and sterilization 2x3 hrsPersonal hygiene-Microbes from hands, tooth-scum and other body parts 2x3 hrs[solation of microorganisms from air, water and soil samples.Dilution and pour plating, colony purification.Enumeration of microorganisms. Total vs. viable counts.Identification of isolated bacteria. Gram staining, other staining methods,

metabolic characterization (e.g. IMVIC test)Growth curve of microorganismsAntibiotic sensitivity of microbes, use of antibiotic discs

Testing of water qualityTest for antibodies against given bacteriaOne step growth of bacteriophageCulture from body fluids (Stool, Urine, Blood)A lcoholic and mixed-acid fermentation

Project: The students will be assigned to generate data on available information from literature

Oil a given topic of biotechnological relevance .

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B.Sc. n J I PaVEl .'1,

METHODS IN MOLECULAR AND .CELL BIOLOGY &ANIMAL CELL CULTURE

UNIT -IMolecular basis of life 02Structure of DNA 01Structure of prokaryotic genes 0 1Structure of eukaryotic genes 02DNA replication both in prokaryotes and eukaryotes 04DNA recombination, molecular mechanisms of prokaryotic and eukaryotic 02insertion elements and transposons 03

I UNIT - IIProkaryotic transcriptionProkaryotic translationProkaryotic gene expression (lac, his, trp, catabolic repression)UNIT - IIIEukaryotic transcriptionEukaryotic translationEukaryotic gene expression in yeast, protozoan parasiteMitochondria and chloroplast and transcription factors etc.Posttranslation regulation of gene expressionDevelopment and environmental regulation of gene expressionUNIT - IVHistory of development of cell culturesThe natural surroundings of animal cellsMetabolic capabilities of animal cellsSimulating natural condition for growing animal cellsImportance of growth factors of the serumPrimary cultures. Anchorage dependence of growthNon-anchorage dependent cellsSecondary cultures. Transformed animal cells - Established/continuouscell lirtesCommonly used animal cell lines - their origin and characteristicsUNIT - VGrowth kinetics of cells in cultureApplications of animal cell culture for studies on gene expressionOrgan cultureTransfection of animal cells: Selectable markers, HAT selection,antibiotic resistance etc.Cell fusionTransplantation of cultured cellsDifferentiation of cells.

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B.Sc.II jl ¥l'\PI=n I( W ~E..~.e."tc...,t.I.MMUNOLOGY & 1i\IMUNOLOGICAL METHODS

UNIT - IThe immune system and immunity along with historical perspective 02Adaptive and innate immunity 01The organs and the cells of the immune system and their function 06Soluble rnediaters of immunity 0 IAntigen-antibody and their structure 03Antigen-antibody interaction 03UNIT- IIHumoral immunity 0 IThe lymphoid system OJAntigen receptor molecules 04Antigen recognition 02Cell co-operation in the antibody response 03The generation of diversity 04UNIT - IIICell mediated immune reactions 03Regulation of tile immune response 02Origin and development of diversity in the immune system 03Complement system 04Cell migration and inflammation 02UNIT - IVEffecter mechanisms 0 IImmunity to infectious diseases 02Invertebrate immunity and vertebrate immunity 05lmmunity to Viruses, Bacteria and Fungi 05Immunity to Protozoa and Worms 04Il11ll1unodeflciency 02UNIT - VVaccine, preparation and types 02IIyperscnsiti\'ity 06

Autoimmunny 0 I

Transplantation 0211lll11unologi~al techniques 06

- Precipitation Reaction in Gels

IlacIllagglutination and Cornp lenn-m Fixation- Direct and Indirect lmmunofluorcsconre- Radioimmunoassay and Enzyme-linked Illllllunosorbcnt Assay (ELISA)

lmrnunob lotting and Immunopreci pi tation- Isolation of pure Antibodics- Assays for complement- Isolation of Lymphocyte populations- Effector Cell Assays.

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(PracticalsPart ICytological preparations:

- Fixation, dehydration and staining- Squash in stain- Embedding and sectioning

Cell counting methods:- The haemocytometer and other aides

Measurements with the help of light microscope- Calibration of ocular micrometer- Finding out average cell size- Chromosome lengths- Separation of cell types (from blood)- Separation of cell organelles- Methods for cell lysis: rupture/osmotic/chemical/enzymatic

lysis of cells followed by centrifugation. Monitoring of cell lysisby release of cellular material and by change in light scattering

etc.

2x3 hrs2x3 hrs3x3 hrs

2x3 hrs

lx3 hrslx3 hrslx3 hrslx3 hrs2x3 hrs2x2 hrs

Extraction of cellular materials- Extraction in saline buffers- Extraction in solvents- Precipitation from extracts- Separation of the constituent molecules of the extraction

in aqueous buffer- Gel filtration- Ion exchange chromatography- Thin Iayer chromatography of ext meted materials- Isolation of chromosomal and plasmid DNA from bacteria- Laking competent Eccoli

- Transfection of plasmid DNA and selection for transformantsPart IIPurification of antigensRClising polyclonal antibodiesPUI ificarion of antibodiesConjugation and labeling of antibodiesEnzyme linked immunoassayRadioimmunoassayRadial immunod iffusio n analysisGt ncrauon of asritic fluidDlagl1o~is of an infectiolls disease by an imrrrunoa s s.,v

Projec t : TIll: ~! Iuk'11i' ',\ ii I be :l:)<.;i~IJL'd \1I ~"·i1'.I'.II, d:ll.1 "11,I'. .11U'k inlouuation Irorn Iiteraturc{In ~l~i\ (on h If ~;\.: •. I ri .it .~ .~\: 1!!-,ph l~.~!\",d rc k:\ . if I, I..

2x3 hrs2x3 hrs2x3 hrs

2x3 hrs2x3 hrs3x3 hrs2x3 hrs4x3 hrs2x3 hrs3x3 hrs

4x3 hrsSpread over 8 weeks

4x3 hrs2x3 hrs3x3 hrs3x3 hrs2x3 hI'S

3x3 hrs

(7\• I

· B.Sc. *1' ]'9% PAIH'I'

ANIMAL CELL BfOTE(~HNOLOGY & PLANT BIOTECIINOLOGY

Unit - IIGrowth factors promoting proliferation of animal cells(EGF, FGF, PDGF, IL-l, IL--2,NGF, erythropoietin etc.) 04Bioreactors for large scale culture of cells 04Transplanting cultured cells 01Introduction to in vitro methods. Terms and definitions. Use of growth 02regulators.Beginning of in vitro cultures in our country (ovary and ovule culture,in vitro pollination and fertilization) 0 1Unit - IIIEmbryo culture, embryo rescue after wide hybridization, and itsapplicationsIntroduction to the processes of embryogenesis andorganogenesis and their practical applications.Clonal multiplication of elite species (Micropropagation) axillary bud,shoot-tip and meristem culture.Haploids and their applications. Somac1onal variations andapplications (Treasure your exceptions)Endosperm culture and production of triploidsPractical applications of tissue and organ culture(summarizing the practical applications of all theabove mentioned techniques).Single-cell suspension cultures and their applications in selectionof variants / mutants with or without mutagen treatment(of haploid cultures preferably).

Unit - IGeneral metabolismSpecial secondary metabolites/products (Insulin, Growth hormoneInterferon, t-plasminogen activator, factor VIII etc.)Expressing cloned proteins in animal cells.Over production and processing of chosen proteinThe need to express in animal cellsProduction of vaccines in animal cellsProduction of monoclonal antibodies

Unit - IVIntroduction to protoplast isolation: principles and applicationsTesting of viability of isolated proroplastsVarious steps in the regeneration of protoplasts

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Somatic hybridization - an introductionVarious methods for fusing protoplasts - Chemical and electricalUse of markers for selection of hybrid cellsPractical applications of somatic hybridization (hybrids vs cybrids]

02010101

Unit-VUse of plant cell, protoplasts and tissue culture for geneticmanipulation of plants. Introduction to A. tumeJaciensTumor formation of planes using A. Tumefacieris (Monocots vs Dicots)Root -formation using A. rhizoqenes/Practical application of genetic transformation.

02010101

RSc.1I1 U"I!t!I HitS 42.."'4..£ rtA....MATHS & COMPUTERS, £1 rYIRONMENTAL BIOCHEMISTRY &

CULTURAL METHODSUnit - IPro babili ty calculati 0nsMethods of sampling, confidence levelMeasurement of central tendenciesMeasurements of deviations.

Unit - II

Computers, General introduction to computers, organisation ofcomputers, digital and analogue computers, computer algorithm.Computers in on line monitoring and automation.Application of computers in co-ordination of solute concentration,pH and temperature etc of a fermenter in operation.

0403

03Unit - HIRenewable and non-renewable resourcesWhat is renewable should be bioassimilable/biodegradableMajor consumer items: Food, fuel and fibresConventional fuels and their environmental impacts: Firewood,Plant and animal wastes,Coal, Gas and Animal oils

)Modern fuels and their environmental impacts:

Methogenic bacteria and biogas.Microbial hydrogen production.

- Conversion of sugars to ethanol. The gasohol experiment.

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Solar energy converters-Hopes from the photosynthetic pigments.Plant based petroleum industry.Cellulose degradation for combustible fuel.

Unit - IVBiotechnological inputs in producing good quality natural fibres-Transgenic sheep and transgenic plantsMicrobiological quality of food and waterTreatment of municipal waste and industrial effluents,

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Unit - V

Degradation of pesticides and other toxic chemicals by microorganisms 02Thuringiensis toxin as a natural pesticide 01Biological con trol of other insects swarming the agricultural fields 02Enrichment of ores by microorganisms 02Biofertilizers. Nitrogen fixing microorganisms enrich the soil with 03assimilable nitrogen.

PracticalsTnitiating plant tissue culture: (dedifferentiation of explants).Growth of plant cells into undifferentiated mass.Large scale cultivation of plant cells in suspension.Induction of differentiation bv modulating the hormonal balanceCulture of lymphocytes from blood samples, preparation of media, filtersterilization, monitoring microbial contamination (bacteria, fungi andmycoplasma) cloning of animal cells by cell and colony. purification.Fusion of cultured cells with myeloma cellsProduction of monoclonal antibodies at a large scaleDemonstration / operation of large scale ferrnenters.

2x34x34x33x3

lOx33x3

Spread over 8 weeks

PROJECTThe students will be assigned to gcnerate data an available information

from literature on a given topic of biotvch nological relevance.

ENTERPRENEURSHIPThe students will be delivered lectures on how to select for a product

line, design and develop processes, economics on material and energy re-quirement, stock the product and rele ase the same for mmketing etc. Thebasic regulations of excise also should be apprise on the candidates. In paral-lel the students will be asked to survey the demand for a given product,feasibility of its production under the givell coristraint s of raw material energyinput financial situations export potential de. Procedural details on how toselect process, how to move for loans, how to operate and how to repay theloans in a phasic manner should also be highlighted during the lectures. Thesemester should end with submission ()( ~1 draft project by trw students.

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