BIOTECHNOLOGY CHAPTER IMPORTANT NOTES, Exams of Biotechnology

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'One - Shot'

@ MIND MAP® D FOR CLASS 12™ Biotechnology Principles and Processes Nn) K: i) t- —— By- Dr. Aarushi Ma’am Topics 44 ..../ ” ee) Biotechnology Principles and Processes ma ‘One - Shot' INTRODUCTION Biotechnology deals with techniques of using|live organisms or enzymes from organisms to produce products and a —— ee —_____. processes useful to humans am Parameters Traditional biotechnology Modern biotechnology Organisms involved Microbes * Genetically modified organisms Production 7 Small scale arge scale Examples/Technique / Curd, bread or “In vitro fertilization leading to include wine making a ‘test -tube’ baby aa Synthesising a gene and using it ° Developing a DNA vaccine ¢ Correcting a defective gene ———_—_————_————__—_— os EFB (European Federation of Biotechnology) The integration of It encompasses natural science and both traditional organisms, cells, view and modern parts thereof, and molecular molecular biotechnology. analogues for products and services’. AR ADVANTAGES OF BIOTECHNOLOGY OVER OTHER ‘= TECHNIQUES Methods |.\Asexual reproductio ll( Sexual reproduction III. Traditional hybridisation IV. Genetic engineering ° Advantage Disadvantage Preserves genetic information. * Novariations 4 * . — Provides opportunities for * Some of which may be harmful to ariations nd formulation of th e organism ae wel asthe unique combinations of genetic population. setup. . Vv on lead to inclust d 2 C é ery often lead to inciusion an ISG x eek Siig atari HIDEO. multiplication of undesirable Allows u us to isolate and introduce genes alon g with desirable genes. only one one or a set 3 set of desirable — genes without introducing undesirable ge genes into target organism +s ~ THREE BASIC STEPS IN GENETICALLY MODIFYING ‘=? ORGANISMS Identification of DNA * Introduction of the identified * Maintenance of introduced with DNA into the host. DNA in the host and transfer ee ee Ea SS A —_—_—_—_—_— desirable genes. tot of the DNA to its progeny. AN} as ot Y CQO PNA ooh E -col wry ser S eat ei ot C leave a) (Fol) GM: of Ving 230 * Restriction endonuclease More thar{909 restriction enzymes have been isolated from strains of bacteria (prokaryotic cell) each of which recognise different rec ition sequences. ————_—— — ee ee __ a + Nomenciature/Naming of enzyme: a + Functions by: at ne ae bn * ‘Inspecting’ the length of DNA sequence y¢ XX XAnome “UVR xX Xx XX * Binds to the “specific recognition sequence” 226 rs — + Cuts the two strands of ds DNA at s pecitic points ’ in their $ and leaves single stranded portions at the ends y OBO SARA DD ee - hese overhanging stretches ond catedeky Ends) DADA | 2 GSD * When source DNA and yector DNA ore cut by the same restriction enzyme the resultant DNA fragments have the same kind of ‘sticky-ends’. Sticky ends are named so because they form hydrogen bonds with their complementary cut counterparts and this stickiness facilitates the action of the enzyme DNA ligase. + First restriction endonuclease Gina) solated and characterised five years later, recognises sequence o(6er. ) Coe. ) + First cecombinant DNA was prepared by Stanley Cohen and Herbert Bo 9725 ——————— + Antibiotic resistant gene a baredunadl + PI id of Sal lla >— Recombinant plasmid 4 Escherichia coh typhimurium Stanley Cohen &% Hetont tose =)4 42. (PY € Gnibiotic aT he Rows tant k Hts joes KZ sions ea ce —(ex2)- E-le CLONING VECTORS Cloning Sites/Restriction Sites Origin of Replication (ori): Selectable Marker: ». Seeuence fone * Helps in selection o Gransformants * Single recognition site for a restriction ‘ aon EES * Normally, the genes encodi a enzyme within the vector is a referable ; 2 stance to antibiotics such as feature. * Responsible f resistance to antibiotics such as Jeature. a Bhp perry ampicillin, chloramphenicol, * Presence of more than one (Se —s ry @y : i —— lo tetracycline or kanamycin, etc., are recognition sites within the vector will ° Those vectors are preferred considered useful selectable markers generate several fragments, which will wnichs upport hi hcopy for E coli complicate the gene cloning ee The normal E.coli cells do not carry * The ligation of alien DNA/gene of number pal AD phate on ant stk dn esl oe 4 : — —— ———— resistance against any of these interest (GOI) is carried out at a antibiotics restriction site present in one of the en — ——_———_—_—O antibiotic resistant genes. SS eee, HL a pens & neon -Recombinan = OC C) Recombinant Shossibili CB — ataale ss aa + If transformation successful - Transformants obtained are of two types: + All transformants are not recombinants but all recombinants are transformants. , es pyene helps in selecting the tronsformonts whereas he other antibiotic resistant gene helps in selection of recombinants * rop—~codes for the proteins involved in the replication of the plasmid | v Non Recombinants Clat Hind Itl EcoR | ul Pst Gamit! E.coli cloning vector pBR322 Pru ll Gene of interest cloned x Resistance toampicillin Vv Resistance to tetracycline ¥ v Recombinants Insertional — inactivation Ls — LQ / amp tet ‘ v v x [due to inactivation] Plasmids as vectors * Extra chromosomal, circular, * Replicate independent of the control * They may have_1 or 2 copies per cell or double stranded DNA of chromosomal DNA (autonomously). even 15 - 100 copies per cell.