Danny Arroyave Zuluaga

Sergio Andrés Romero SeguroTERCER SEMESTREPROFESORA: LINA MARIA MARTINEZ

Is a multisystem disorder with an autosomic

dominant pattern.

20–89% of the cases to mutations in the JAG1

gene.

JAG1 gene is located at 20p12.2.

Presents so far more than 300 mutations.

ALGS has been one of the main reasons for

referral for transplantation of liver.

organ alteration

Liver particularly presenting bile duct paucity

heart peripheral pulmonary artery stenosis

kidneys renal dysplasia, renal tubular acidosis, among others

skeletal system butterfly vertebrae

characteristic facialfeatures

broad forehead, deep-set eyes, pointed chin and a triangular face

eyes

Polymorphism

alternatives are isoforms of a gene in a population. affects both gonadal and somatic cells, so it can be transmitted following the laws of Mendelian inheritance.

Mutations

permanent change occurred in the base sequence of the DNA of an organism. are only affects inheritable when gonadal cells and not when exclusively affects somatic cells.

recognize and investigate the JAG1 gene, which could help us in the clinical monitoring

of these patients.

Extracción de Dna:

Ruptura de las celulas (detergentes y enzimas).

Eliminacion de proteinas.

Eliminacion de ARN.

Extracción de proteinas.

Precipitaciones del ADN.

DHPLC

Técnica cromatografía para la separación y el análisis de fragmentos de ADN con diferente longitud y/o composición de base

RT-PCR

Amplificación de RNA a través de la síntesis previa de su DNAc (copia), utilizando una transcriptasa inversa, seguido de varios ciclos de PCR convencional. Con esta técnica se puede determinar la expresión de genes en diversos tejidos.

Secuenciación Tiene métodos tanto químicos como

enzimáticos

a) Químicos: marcaje del DNA, hidrólisis química selectiva del DNA y análisis de los productos

b) Enzimático: síntesis enzimática, análisis de los fragmentos y automatización

Su objetico es la amplificación directa de un gen o un fragmento de DNA o indirecta de un RNA

Se basa en amplificación enzimática in vitro, que es el incremento geométrico del numero de copias de una secuencia particular del DNA

Técnica metodológica

1. Desnaturalización: calentamiento para las separación de las dos hebras de DNA, mediantes incubación breve a una temperatura entre 90-97 grados centígrados

2. Alineamiento: hibridación de las hebras sencillas del DNA de interés con los primers y DNTp's. A una temperatura de 37 a 65 grados centígrados

3. Síntesis: amplificación en la que la DNA polimerasa elonga los primers empleando como molde ambas hebras originales. La replicación transcurre en dirección 5‘ a 3‘.

Fig 1. A y C NM por DHPLC ; B y D Electropherogram.

Fig 2. Mutación por splicing Nuevo con especificación entre La región exon1 a exon2.

Fig 3. Perdida de exón 4 confirmada Posterior a RT PCR.

AUTHOR WHAT DID HE SAY? THEY ARE AGREE? YES OR NOT

(Onouchi et al., 1999)

“In the case of the intron 4–5 mutation described (and as has been previously reported for other abnormal splicing mutation affecting the following downstream base in the donor splicing site), thischange could generate a less severe phenotype.”

NO

Heritage et al. (2002).

“The present study further supports the idea that the molecular mechanisms involved in the clinicaleffect of the mutations in ALGS is the haploinsufficiency mainly due to the loss of the transmembranal orthe DSL domains (NMD).

YES

AUTHOR WHAT DID HE SAY? THEY ARE AGREE?YES OR NOT

(Kamath et al., 2012; McDaniel et al., 2006).

“Finally, in our study we observed that in 2 patients with ALGS clinical diagnosis there were nomutations identified in JAG1, however, we cannot discard the presence of mutations in other regions of thegene (such as the promoter) or in NOTCH2 gene”

YES

(Mátyás et al., 2002).

“It has been demonstrated that polymorphic changes detection is difficult when there are two variations inthe same amplicon”

YES

1. Is really important that we (medicine society), start a process in the early determination of mutations in the newborn.

2. The field of the mutations moves really fast, and maybe the solution is not try to stop it, is work hard and fast as they do.

3. We think that it could be a good solution, the implemetation of a law that allow the people to make a genomic exam for free.

4. The people should know more about its history, and where they are from, this would be really helpful to make a geographic location and try to know, when all these things start and where come from.