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ANLISIS DE

RESTRICCIN

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1970

Base Qumico usado parala alteracin de la

base

Qumico usado pararemover la base

alterada

Qumico usadopara dividir la

cadena

G Dimetil sulfato Formato de piperidina Piperidina

A + G Acido Acido Piperidina

C + T Hidrazina Piperidina Piperidina

C Hidrazina + alcali Piperidina Piperidina

http://localhost/var/www/apps/conversion/tmp/scratch_7//commons.wikimedia.org/wiki/File:Guanine_chemical_structure.png?uselang=es

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Haemophilus influenzae

HindII

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El Premio Nobel de Medicina de 1978 fue concedido a losmicrobilogos Werner Arber, Daniel Nathans y HamiltonSmith por el descubrimiento de las endonucleasas de

restriccin lo que condujo al desarrollo de la tecnologa deADN recombinante. El primer uso prctico de su trabajo fue

la manipulacin de la bacteriaE. coli

para producir insulinahumana para los diabticos.

http://localhost/var/www/apps/conversion/tmp/scratch_7//upload.wikimedia.org/wikipedia/commons/9/9a/NobelP2.png

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Nomenclatura Ejemplo Corresponde a:

E Escherichia Gnero de la bacteria

co coli Especie de la bacteria

R RY13 Cepa de la bacteria

ILa primera enzima

identificada

Orden de identificacinde la enzima en la

bacteria

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Extremos compatibles

Isoesquizmero

http://upload.wikimedia.org/wikipedia/commons/e/ea/Isoesquiz%C3%B3meros.pnghttp://upload.wikimedia.org/wikipedia/commons/e/ea/Isoesquiz%C3%B3meros.png

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Endonucleasa Metiltransferasa

S-adenosil-metionina(ATP+metionina)

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SISTEMA DE RESTRICCION MODIFICACION TIPO I

SISTEMA DE RESTRICCIONMODIFICACION TIPO II

ENZIMA TIPO I ENZIMA TIPO III ENZIMA TIPO II

Enzima bifuncional de 2subunidades: R y M.Acta simultneamente.

Enzima bifuncional de 2subunidades.Endonucleasa y metilasa.Acta exclusivamente

Endonucleasa. Actan porseparado.

Sitio de reconocimiento:5-7 pb. 5-7 pb. 4-6 pb.

Corta al azar.100-1000 pb.

Corta al azar.24 - 26 pb.

Cerca del sitio dereconocimiento.

Cohesivos. Cohesivos. Romos o cohesivos.

ATP, Mg y S-adenosil-metionina.

Mg, ATP y SAM. Mg y SAM

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1) Marcador de 1Kb.2) Control negativo.3) Digerido conApa IySal I.4) Digerido conApa IySal I.5) Digerido conSal I.

Apa I Sal I

Sal I778Sal I154

pGEM-Tpsr4182 pb

pGEM-Tpsr(4182 pb)

Apa IySal I 3000, 624, 408 y 154 pb.

Sal I 3154, 624 y 408 pb.

506, 517

1 2 3 4

396

3054

3000 3000

3154

624 624 624

408 408

5

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Problema 1. Qu individuos tienen un

alelo normal y qu individuos uno

mutado? Justifica tu respuesta

3 kb

0.75 kb

0.5 kb

Problema 2. En la fotografa se muestran 3

plsmidos con inserto, podras decirme si

son iguales? Justifica tu respuesta

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Glick, B.R., Pasternak, J.J., Molecular Biotechnology: Principles and applications of

recombinant DNA, Ed. American Society Microbiology, 1998.

Sambrook, J. y Russell, D.W. Molecular Cloning: A laboratory manual. Ed. Cold Sping

Harbor Laboratory Press, New York, 2001.

http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/default.asp

(NewLab)

http://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-

resource (Promega)

http://www.fermentas.com/en/products/all/fastdigest-restriction-enzymes

(Fermentas)

http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/default.asphttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.fermentas.com/en/products/all/fastdigest-restriction-enzymeshttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.promega.com/resources/product-guides-and-selectors/restriction-enzyme-resourcehttp://www.neb.com/nebecomm/tech_reference/restriction_enzymes/default.asphttp://www.neb.com/nebecomm/tech_reference/restriction_enzymes/default.asp