Documenti di Didattica
Documenti di Professioni
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Purinas:
Pirimidinas:
terminal 5'
HO O
Adenina A
NH
2
O O P HO O O N
N
O
Citosina C
O N
O P HO O O N N
NH
NH
Guanina G
O CH O P HO O O N
3
NH O
Timina T
OH
terminal 3'
LA INFORMACION NECESARIA PARA GENERAR LAS PROTEINAS INCUYENDO LAS ENZIMAS SON CODIFICADAS POR GENES.
Expresin gentica
promotor Secuencia de unin al ribosoma Gene estructural (secuencia codificante) codn de inicio codn de paro terminador
ADN
P RBS
Transcripcin
ARN
RBS Traduccin
Protena
Dogma Central
Transcripcin
Synthesis of RNA from a DNA Template. Requires DNA-dependent RNA polymerase plus the four nucleotides (ATP, GTP. CTP and UTP). Synthesis begins at a the initiation site on DNA. The template strand is read 3' to 5' and the mRNA is synthesized 5' to 3.
Transcripcin
Transcripcin
Cdigo Gentico
Traduccin
Herramientas
Enzimas de restriccin (endonucleasas)
Enzimas de modificacin Ligasa ADN polimerasa Nucleasas kinasa, transferasa,etc,etc,etc. Vectores (plsmidos, fagos, csmidos, etc)
AISLAMIENTO DE UN GEN
http://nobelprize.org/chemistry/educational/pcr/
Enzimas de Restriccin
Enzimas de Restriccin
BfuI
BciVI
GTATCC(N)6/5^ GCCNNNN^NGGC A^GATCT CC^NGG GACNN^NNGTC GAAGAC(N)2/6^ ^8/13(N)GAG(N)5CTC(N)13/8^ CCTNAGC-5/-2^ GC^TNAGC C^CNNGG
ER1501/2 ER0071/2 ER0081/2 ER1421/2 ER1431/2 ER1011/2 ER1311/2 ER1181/2 ER0091/2 ER1081 ER0871/2 ER1711 ER1201/2 ER1261/2 ER1401/2 ER0881/2 ER1441/2 ER1451/2 ER0921/2 ER0891 ER1001/2 ER1461/2 ER0111/2 ER0121 ER0131/2/3 ER0781/2 ER0791/2 ER0931/2 ER1151/2 ER1321/2 ER0831/2 ER0701/2 ER1021/2 ER0141/2 ER0151/2/3 ER1741
Enzimas de Restriccin
BoxI BpiI PshAI BbvII
Fermentas Specificity Prototype Enzyme 5'=>3' AarI AarI AasI DrdI AatII Acc65I AdeI AlfI AloI AluI Alw21I Alw26I Alw44I ApaI new BamHI BauI BclI BcnI BcuI BfiI BfmI BfuI AatII KpnI (GGTAC^C) DraIII AlfI AloI AluI HgiAI BsmAI ApaLI ApaI BamHI BsiI BclI CauII SpeI BfiI SfeI BciVI
Catalog # CACCTGC(N)4/8^ GACNNNN^NNGTC GACGT^C G^GTACC CACNNN^GTG ^(10/12)GCA(N)6TGC(12/10)^ AG^CT GWGCW^C GTCTC(N)1/5^ G^TGCAC GGGCC^C G^GATCC CACGAG(-5/-1)^ T^GATCA CC^SGG A^CTAGT ACTGGG(N)5/4^ C^TRYAG GTATCC(N)6/5^ GCCNNNN^NGGC A^GATCT CC^NGG GACNN^NNGTC GAAGAC(N)2/6^ ^8/13(N)GAG(N)5CTC(N)13/8^ CCTNAGC-5/-2^ GC^TNAGC C^CNNGG ER1581/2 ER1721/2 ER0991/2 ER0901/2 ER1231/2 ER1801 ER0011/2 ER0021/2 ER0031/2 ER0041/2 ER1411/2 ER0051/2/3 ER1841 ER0721/2 ER0061/2 ER1251/2 ER1591/2 ER1161/2 ER1501/2 ER0071/2 ER0081/2 ER1421/2 ER1431/2 ER1011/2 ER1311/2 ER1181/2new ER0091/2 ER1081 ER0871/2 ER1711
BplI Bpu10I
Bpu1102I BseDI BseGI BseJI BseLI BseMI BseMII BseNI BseSI BseXI Bsh1236I Bsh1285I BshNI BshTI Bsp68I Bsp119I Bsp120I Bsp143I Bsp143II Bsp1407I BspLI BspPI BspTI Bst1107I BstXI Bsu15I BsuRI BveI
FokI (GGATG(N)9/13^) GGATG(N)2/0^ BsaBI GATNN^NNATC BsiYI CCNNNNN^NNGG BsrDI BseMII BsrI BseSI BbvI FnuDII McrI HgiCI AgeI NruI AsuII ApaI (GGGCC^C) MboI HaeII Bsp1407I NlaIV BinI AflII SnaI BstXI ClaI HaeIII BspMI GCAATG(N)2/0^ CTCAG(N)10/8^ ACTGG(N)1/-1^ GKGCM^C GCAGC(N)8/12^ CG^CG CGRY^CG G^GYRCC A^CCGGT TCG^CGA TT^CGAA G^GGCCC ^GATC RGCGC^Y T^GTACA GGN^NCC GGATC(N)4/5^ C^TTAAG GTA^TAC CCANNNNN^NTGG AT^CGAT GG^CC ACCTGC(N)4/8^
new
^7/12-13(N)GAAC(N)6TCC(N)12-13/7^ ER1491/2
BglI BglI BglII BglII Bme1390I ScrFI BoxI BpiI BplI Bpu10I Bpu1102I BseDI BseGI BseJI PshAI BbvII BplI Bpu10I EspI SecI
Ligacin
T4 DNA ligasa
VECTORES O PLASMIDOS Son molculas de AND Con origen de replicacin. Muchas portan genes de resistencia a antibiticos. Pueden transferirse de una clula a otra.
VECTOR DE CLONACION
Vectores
4363 pb
2686 pb
Vectores de expresin
PROMOTORES Inducidos por diversos factores.
Inductores: Calor Fuente de carbono Presencia de algun compuesto inductor
SELECCION
Opern lac
CLONACION DE UN GEN
DIVERSA
Aisla enzimas de fuentes naturales Obtiene muestras de diferentes regiones del mundo. Aisla el ADN (metagenoma) Construye bibliotecas genomicas Caracteriza con equipos roboticos miles de clonas a la vez, tanto sus capacidades cataliticas como sus propiedades cineticas
Expresin heterloga
ENZIMAS
INGENIERIA DE PROTEINAS
Recombinant Chymosin
PROTEINAS DE LA LECHE
Chymosin Reaction
Bacillus sp. Bacillus species are non-pathogenic and are used industrially to produce several enzymes used in food processing such as amylases. B. licheniformis could produce the chymosin, but the signal sequences did not allow the secretion of the enzyme into the medium. by addition of benzoic acid and the chymosin is isolated by filtration.