Class Note for MIC 205A at UA 4
Class Note for MIC 205A at UA 4
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Date Created: 02/06/15
Biotechnology and Recombinant DNA WERL39S A We TENN or DNA Roweme BEND cum Nazi s A sum aura esumcmmon THE Thuks ka 9111 3155 601 TD 85 A DREW Biotechnology The use of microorganisms cells or cell components to make a product Foods Vaccines Antibiotics Vitamins Biodegradation Selective breeding Genetic Engineering Manipulating an organism s genome to alter microbes plants and animals for our benefit correct genetic defects in humans Other Uses of Genetic Technology Genetic technologies also used to identify individuals UA part of Shoah Project to ID Holocaust victims Wildcat October 4 2006 UA project to trace ancestry Diagnosis and epidemiology of disease Useful Properties of DNA DNA sequences specify gene locations and protein amino acid sequence Restriction endonucleases cut at specific nucleotides size of pieces gives us information about DNA sequence Nucleotides hydrogen bond with complementary nucleotides DNA hybridization allows recognition of specific genes A probe is a piece of complementary DNA of known sequence labeled with radioactivity so it can be detected DNA Fingerprinting Forensics A t a 7 x l i la 1 squot L 7 J1 rl39 Samples DNA la Cells lrom dillerent samples are FORENSIC TEST if e a i iv A i C39 LE 7 quot439 2 KQ Low mpg N POSItID oi Migration Visualized Bands E A n M w z lragments near the wells smaller ones tanner lrom the wells The relative positions 0 lhese lragmenis are made visible by labeled DNA probes designed to attach to specilic DN markers The to the sample39s pattern N Suspec tz U w Suspectt I I I O I I m Marker 000000 0 o o I v Evrciencez I 39 l m Evrdence1 m Victim Om u Marker OOIOII o lae1 iRanrlQ E S n s vidence Sam les uspecls 1 and 2 suspect lanes which indiVidual committed the rapequot DNA Fingerprinting Epidemiology E caliisolates trorn I patients who drank lt E coli isolates from contaminated juice O m p a S O n patients whose infections we ltApple juice DNA from a I 39 in u a n I l39ll39 p not juice relatfdgt A if fOOd 5tiuiinilu 39iquotii39 s as zsiglz Illgl a Ioliisli bogp39p39 t Useful The compleme Properties of DNA ntary strands of DNA can be separated and associated by heating and cooling One strand of DNA specifies the sequence of the other strand mRNA specifies the sequence of gene DNA re I nMi L 39 4 quot nMA the S 1 39 39 39 4 When cooled the two strands rejoin at complementary sites The two strands 1 Polymerase Chain Reaction PCR Amplifies DNA Sawing 11 Tie McGuallill CLI l39i39Jul i ua llJ l 55 named lLii iupiuuu mi or it w m Inc ie Il rw v r ii 1 c a V quotI pmmg 3w gt avhcv temperature Yhe M rusuttiiiq sl39nnix r39teri semis as rri39iriales lor a 3900 Cyril ct denaturaisn pii39ririq vIHE M vi 39 Cyde I one Sample 3v st 3924 gtaLLLLLLELL eLLLLLLLLquoty AdiJ V Heal to 2139 C r iL E i ri 3 m w a 3 Slrdlih 5 is L39 39f t 444 i4 Wm heartburnbumhabitually v43 39 so lD Gii c V W V I LL3Li3 39 V WC A iicoi is mm P iiiii r i 3 39u MauiAmenity w rim 3 39 39 39 l l l I J w Jal kaLnkLghkpilihlLum tsu39 Primers specify what DNA is copied PCR Amplifies DNA b A View at the process after 6 cycles w39 copies 0 amp ilied DNA Continuing this process for 20 to 40 cycles can produce millions ol identical DNA molecules iuweif h i i ammslliii i mwmsMNWWMNWWWlMR Diagnosis Epidemiology lf EE 7 fEJH Ji 4 copffj L ll Jig xv xquot a Beanies J it it ll ll it it ll i i irlr i ir i inir i ll iliiilililiiliiili iWhhMWmi 39 l39llilkll39l l39 i l39 i lu JUUUUUUUUUU UUU l JUUUUUUl UUUUUl i Ii i ii iiquotlquotiiquot i4 tliini I i w Genetic engineering Recombinant DNA bacterium chromosomes 390 3 o l lplasmcs F isolated human cell 1 Jll ifquot plasm a Combining DNA from 32131 sliced up39 Em enzyme Ch omosome f quota a gene inserted gene responsible for k z controlling formation ofinstiiin 39 7 l ev E h 1 in olasmid t o different 0quot mo bacterium quotquot wasquot plasmid with gene 0 a n I S m S I 39 inserted in bacterium reprocuction an quot ii of quot F T 395 i bacteria 3503 0 33 plasm ds ng 7quot 39 o r 003 03 r kegquot 1quot o t 39339 14 ffquot 335 m 7 prcduction igw 39 of if lSUiquotl Selection of Altered Cells Useful Properties of DNA Culture at cloning host after incubation wi h recam nan tl Tt w mu r p r J t a r Bacteria wrtti manna t r tr mumcram is 39 recombinant Restriction endonuclease Bacteria lack lasrnid V 39 Restriction P r w W endonucleases i i Z GATE 2 if n D Eactgria carrying Ampicillin lasrnid resistance gene 7 39 I u r GALQ f t V 7 75 quot 3quot as S I C S I Regular nonselective Selective medium 2 my Ime gun Wl yvo with ampicillin Orga nism yf Wk N ype a Coanles i Pure culture r otbacteria containing Cloned gene Organism 2 ends for Insertion 8995 HA Se recognizes and cleaves DNA at the site at a specilic palindromic sequence Cleavage can produce staggered rails called sticky ends mai accept 39 39 39 39 complements rat is gas sprung 2 The My and be its 0 0 An tl b I 0 C re SI sta n Ge 3 n e u sed to Join DNA tram ditlererrt organisms by Cutting It with the same restnchon enzyme ensuring tna all lragments have complementary ends 1Action or restriction endonucle restriction endanuclea r Trastuzumab i H EPTIN With Diluem m1 50 P H d 6 Genetically Modified i 7 T T l Organisms Alphainterieron Therapy for leukemia melanoma and hepatitis produced by E coliand Saccharomyces cerevisiae yeast Antitrypsin Assists emphysema patients produced lay genetically modi ed sheep H e I d e I p I a n Betainterferon Treatment for multiple scierasis produced by mammalian cell culture I Bone morphogenic proteins Induces new bone formation useful in healing tractures and reconstructive surgery n g e n e m produced by mammalian cell cullure Colonystimulating Factor CSF Counteracts etiects of chemotherapy improves resistance to infectious disease such I as AIDS treatment oi leukemia produced by E coil and S cerevisiae Epidermal growth factor EGF Heals wounds burns ulcers produced by E coli in Erythropoielin EPO Treatment of anemia produced by mammalian cell culture Factor VIII Treatment at hemophilia improves clotting produced by mammalian cell culture F I S m Gammainterteran Treatment at chronic granulamatous disease produced by E coli Hepatitis B vaccine Produced by S cerevisiae that carries hepatitisvirus gene on a plasmid G I t t C Human growth hormone thGH Corrects growth deficiencies in children produced by E coli 0 e n e a e n 6 Human insulin Therapy Tor diabetes better tolerated than insulin extracted from animats produced Plantbased vaccines by Escherichia coli39 Cayman 39 3304 Puriin Edilwllnn n pic stilnq as Eg39vjmln Cumminqs Transgenic Animals Knockout and transgenic mice used to study immune system and genetic diseases Pigs blood clotting Factor VIII organs for transplantation Others Human lL2 cancer albumin blood volume growth hormone tPA dissolves clots Genetically Engineering Humans Bone marrow supplies stem cells Successful replacement of gene for enzyme needed for lymphocyte development Difficulties in Genetically Engineering Humans Inserting gene in correct cells Inserting gene so it is expressed correctly Orientation Regulation Controlling virus vector Ethical issues Other Issues How will genetically engineered organisms affect environment Spread of genes to other organisms Who will decide We39ve genellcally engineered a tuna exactly the same diameter as our cansquot
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