7/28/2019 Anlisis de restriccin
1/16
ANLISIS DE
RESTRICCIN
7/28/2019 Anlisis de restriccin
2/16
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=es7/28/2019 Anlisis de restriccin
3/16
Haemophilus influenzae
HindII
7/28/2019 Anlisis de restriccin
4/16
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.png7/28/2019 Anlisis de restriccin
5/16
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
7/28/2019 Anlisis de restriccin
6/16
7/28/2019 Anlisis de restriccin
7/16
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.png7/28/2019 Anlisis de restriccin
8/16
Endonucleasa Metiltransferasa
S-adenosil-metionina(ATP+metionina)
7/28/2019 Anlisis de restriccin
9/16
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
7/28/2019 Anlisis de restriccin
10/16
7/28/2019 Anlisis de restriccin
11/16
7/28/2019 Anlisis de restriccin
12/16
7/28/2019 Anlisis de restriccin
13/16
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
7/28/2019 Anlisis de restriccin
14/16
7/28/2019 Anlisis de restriccin
15/16
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
7/28/2019 Anlisis de restriccin
16/16
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.aspTop Related