TP6 : Les UV, un exemple d’agent mutagène

Levures exposées aux UVPrérequis :

Une mutation est un phénomène rare qui correspond à un changement aléatoire dans la séquence des nucléotides d’un gène.

Des agents mutagènes peuvent considérablement augmenter la fréquence des mutations.

Les UV sont des agents mutagènes responsables d’un nombre croissant de cancers chez l’homme.

Les levures sont des champignons unicellulaires qui se reproduisent par bourgeonnement. Chaque levure, en se multipliant, donne naissance à une colonie qui devient visible à l’œil nu lorsqu’elle est suffisamment développée.

Levures exposées aux UV vues au microscope électronique :

[important]

Consignes :

  • Concevoir un protocole expérimental pour tester l’effet mutagène des UV.

Le matériel mis à disposition est le suivant :

– des levures rouges,

– 4 boîtes de pétri contenant un milieu de culture complet (mélange d’acides aminés, glucose, agar, eau distillée),

– une boîte à UV.

Il faut penser à :

  • Indiquer ce qui varie entre les différentes du milieu de culture.
  • Repérer la boîte témoin et indiquer son rôle.
  • Préciser les raisons (au moins) pour lesquelles il faut bien étaler les cellules.
  • Remarque : Le milieu de culture est un milieu complet (mélange d’acides aminés, glucose, agar, eau distillée). Expliquer pourquoi une telle composition est obligatoire pour cultiver des levures.

La semaine suivante :

  • Soit avec vos résultats (s’ils sont significatifs ou de bonne qualité) ou avec l’image disponible sur le réseau « résultats_levuresUV.jpg« , compter les colonies (possibilité d’utiliser Mesurim) et compléter le fichier openCalc « TP6_levures_UV.ods« .
  • Avec le même fichier OpenCalc, réaliser un graphique montrant la relation entre le pourcentage de colonies blanches et du nombre total de colonies en fonction du temps d’exposition aux UV.
  • En utilisant les résultats, les informations extraites des documents suivants (avec Anagène) et vos connaissances, vous devez déterminer : le génotype et le phénotype moléculaire et cellulaire de la mutation, montrer en quoi les UV sont des agents mutagènes.

[/important]

[notice]

Fichier pour le TP :

[/notice]

Document 1 : Gènes et couleur des levures

  • Les levures utilisées sont de couleur rouge.

Cette couleur est due à un gène, le gène Ade2 qui est impliqué dans la chaîne de biosynthèse de l’adénine. Cette chaîne de réactions comporte 12 réactions chimiques successives. Le gène Ade2 contrôle la réalisation de la 6è réaction.

Quand le gène Ade2 est défectueux ou non fonctionnel, la totalité des réactions de la chaîne de biosynthèse ne se déroule plus, la 6è réaction n’a pas lieu et il y accumulation dans le cytoplasme de la levure de la molécule 5, qui en s’oxydant donne une teinte rouge.

  •  Les levures sauvages sont blanches.

Document 2 : Séquence du gène Ade2

Le gène ade2 est le gène responsable de la coloration des levures. Le fichier d’Anagène ade2.edi contient les deux versions de ce gène. L’un correspond à une levure rouge (ademoins) et l’autre à une levure blanche (adeplus).

 

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