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| Date / if a] [DOM rage no 1 a Biotechnology : Principles and Processes g4 She techniques of using five organisms or enzymes fram organisms —_. ———fo_produce products and Processes useful fo humans. (any processes _- like in vitro fertilization ead ing fo “lest -tube’ baby , synthesizing —___| gene and using it ,-develaping a OWA vaccine or correcting a defectiv —_———gene_are_also_ parts_of Biotechnology « ————————= lll =— The European Federation of Biotechnology (EF) Aas given a defini for ) -of biotechnology that comprises both traditionc and madern molecule | biotechnology + The defination is_as follows - i; . ae — m ~— allow us to isolate and fntraduce anly one of a— set_of desirable == genes without introducing undesirable genes _into. the farget orp Alte, i 4 _In_a_chromasome there ts -a_specittc _ DNA sequence called fhe Origin, di of replication to hich is__responsible for initiating replications Therefor, _ _for fhe_multiplication of dny alien piece of .DWA_in-an_otganism * it needs to be a part of a chromasame—whi 4, £ «ce P known as Sorigin of replication’, Thus, an alfen DwA fs linked sith P the origin of replication, so that , this alien piece of DNA can replicate F -and_tmultiply itself in the host organism» This isknown_as©lonins ; / e@ oa - ie ing out a piece of DIVA - a Py ° rs} J oreo « °o Q f ‘ o 9 us “ :. ; 7 ° ((tdeiegi- ¢ NY plasmic ol Ado o ofeye n > C6 Mn ¢ : ° o A © DNA Info fh ale organist é ° Fe) os * ye @ ¢ é J ALA tia e768. LE of Sak a avaeé Ow d t i a a ad rad 2 ay q a 4 jis él¢ 0 J 4° i é b » ¢ 4 , ? a 4 es o Tole l 2S KUN 01fF1 5 Jit an* lf WIOKRES 7 fh IMD iN ¢ jal ee mtionting “DONA. created Yr Wien Aaa bee pe a — Se ; oe a : Example, the first restriction endonucleases — Hind. TL,always cut _ DNA molecules at a_particular point by recognising a. specitic = sequence af six base pairs. This specific pase sequence [s_bnovan ese 1 (3 the Recognition fequence far _Hind ff. __—_————__—— ae : Lach_restriction endonuclease recagnises_a_spectfic_palindramie __ ~___ tucleotide sequence fn the DIVA 2 Palindromes—are graup oF {etters - that form the same word when read bath forward and backcard, ~ | For example , ““MALAYALAt ””. ——— a _§’? — GAATTC — 3? = ——s 4 | 3? —— CTTAAG— S” __ ~______[he_palindrame in DNA is a sequence of base _pairs that reads same ~ on two 5 frands when arfentation sf reading ts kept same. ——-— 7 The enzyme cuts both DNA EcoRI cuts the DNA between bases strands at the same site G and A only when the sequence = Ps | GAATTC is present in the DNA i 4 Vector DNA p Foreign DNA = ’ = \\ fj \\ T G i ct cfr fr A fal : a ( | p a | EcoRI “2 q Sticky end 7 TOO Giza \ TO OWA T iT) A “a | G ——T Sticky end — —— DNA fragments join at sticky ends = ’ | ! y :" | — a | — era | Recombinant DNA | _ __| Restetetinn enzymes cut the strand of DNA a Tite quay fram_the = ; _Céntre of the £2 indrome site between ihe same fwo bases on the —+oppasite strands having sticky strand . The Stickiness of the strands — —_____._ facilitates the actian of the enzyme DNA_d& Ligase. —— -_—_———- ee —- — ——— ee se | Restriction endonucleases are-used In qenatte. engineering ta form —_ ____ ferombi a ee of DONA from : _ied Cut fc ti roe a a : the 4 , Ay z e.4 s = DNA Si = | {gases = Forcign DNA 3000000000000 C8 Same restriction enzyme cutting both foreign ; DNA and vector DNA at specific point ie, § Ligases join foreign - _ DNA to plasmid eo é. _._-—- Diagrammatic Recombinant DNA representation Molecule J transformation S| Page to, 7 Date / / an — mi antibiotics suchas ampicillin, chloramphenicol, tetracycline ar hanamycin : etce, are considered! useful selectable markers for Eo colf. : ——a Cs Cloning sites + 5 es: a el fa: link the foreign. DIVA, the vector need fo have single recagnitéon ———Sites_for the commanly used restriction eniymes as presence of more — _than_one tecagnition. sites within the vector will generate several fragments, which will complicate the _gene cloning» o The_ ee of : fade BOLRte abies at aig in_ane of the - Pvu fl eo? o , ° if TiC uv on stele ¥ Jeitale , ry a “lai #] hed ehaatela f) o -* ACSIMTIIG t py? e ela e ° f ad ote f ug ° g 4 . olelee ' e : 1] A q - j07ino ¢ r L ] Ceri {; aes ‘ i Pse)' ened ’ afte eneig é a one é ‘4 arleve Py r " . f) jdesio tf / ‘sa gGelga PPE mT x £488 ° “ _ = e mdtele a ite : : ‘ » | Wale al onete vale s ie @ P " a a 10) 7 a — i 7 flen or TJ all re Velc - No jt Jere €fic oc * : ae 5 a { i. = a = = : 7 “ H ‘. _ _beta- galactosidase, which can cleave a chramogenic substrate ane a_blue coloured product olf this lac Z gene is inactivated by = Jnsertion_of a target. DNA fragment into it, the development Of the | blue colour wilt be prevented and it gives white coloured colonies. _ By this way ,w e can differentiate recombthant Cuhite colour) and | nan- recombinant (blue calour) colon (Cs a =... i fs and animals: [ aciens (pathogen of dicat plant) is_able fo deli 6 T- »” fa transform normal —____1 plant cells into a tumor ahd clfrect these tumor cells fo produce ______| the chemicals required by the pathogens «Retroviruses in_antmals — al A See : [ { ( L ( [ has ___| been modified into cloning vector having no more pathogent. fo — _ ( ot r/ | ( c h | (EE; | a f ( 6 le to} * ° , ‘° a ° e ° J Ss 1 ang) NeMMice at men n civaien t oa 0 a ; 2 a a ¢ nue didelétele @ ele ? ' ry i (| f] ° Fy NA plasmic 2 © ° DNA qn @iso 0 qansformeca nfo ne f, 5 A : ° alse P) ale Nem DPpacr OF! 2 a eterna at aay ote ¢ é a é y th combinant DIVA o - 2 * oa 3) 2 bow bl 8 ola igiacetels U INA af a 4 ILC a e oof a ’ i th ictéimé vate ia oneyee ’ 4) EL i 1 - f é a LL LOL it r) a4 wold rf ff “ie oy 4 a JNA inte nath esueta S 4 v4 Ah =4 @ aeinat LT —— Rn Amplititation of (nene of Interest using f — Ta get multiple copies of fhe D et of primers and enryme DNA. f —using_s Ui ul “ Region to be amplified UR % DNA polymerase (Tay polymerase) + deoxynucleotides Amplified {(~1 billion times) (YA_or. gene of interest tn vitro by olymerase« | | WOM | Page No {[ : | Date / / - ——-+ ————_———————————— [nsertion of Recombinant ONA into the Host Cn / Organism 3 ___ It includes making the recipient cells competent fo receive, take up. —________| DNA present in. ifs surrounding, ¢tcs The recombinant. DNA bearing __ gene for resistance fo an antibiotic fs transferred fnto Ecol’ cells , el ______the hast cell become _transfarmed_ into ampicillin -resistance cellg5 $$$ Ob ign ne_neaduct 4 j BE aR: a ee _dnimal cell fo taprduce destottle Proteins Expression af foreign genes in host Cells nena ce Condition ta obtain tecombinant pratein «The recombinant cell is = : | 134 Tt ° fi é (f - f (1 rhe y { mn 7 1 '§ : _____drained out from one side whi jum_ if e ‘ ° ” © © ] = — Increased surface —— ~ @area for r Acid/Base — an for pH iz control : braker = e |. seein PME Patent | sterilisation 1% pas es a , “> Bubbles\ __ dramatically increase the — oxygen ~ Sparged stirred-tank bioreactor through which = sterile air bubbles are sparged — 
