Paper Information

Title: 

DESIGNED A NOVEL RECOMBINANT VECTOR THAT CAN INSERT SUICIDE GENES, HSV-TK & YEAST-CD INTO THE SSU RRNA OF LEISHMANIA GENOME BY HOMOLOGOUS RECOMBINATION

Type: SPEECH
Author(s): DAVOUDI N.,DOUSTI A.
 
 
 
Name of Seminar: IRANIAN CONGRESS OF PHYSIOLOGY AND PHARMACOLOGY
Type of Seminar:  CONGRESS
Sponsor:  PHYSIOLOGY AND PHARMACOLOGY SOCIETY, MASHHAD UNIVERSITY OF MEDICAL SCIENCE
Date:  2007Volume 18
 
 
Abstract: 

Introduction: Leishmaniasis is a widespread, infectious parasitic disease Caused by Leishmania spp. There is no vaccine against any form of leishmaniasis for general human use. Leishmania can be genetically modified and used as a vaccine. The aim of this study was designed a construct to insert "suicide" genes; HSV-tk and Yeast-cd into small subunit (SSU) rRNA of Leishmania genome by homologous recombination.
Methods: In order to provide the tub gene fragments were cloned into a construct, at the first step tk, cd and tub-cd arrangement. The fragment containing;
pBluescript vector and make the tk- tub-cd from pBluescript vector was digested by SalI/NotI and subcloned into atk- pF4x1.4sat vector by stuffer fragment replacement. pF4X1.4sat vector allows directional insertion of the gene fragments into the chromosomal 18S rRNA locus ssu making use of the high level transcription rates.
Resulted: The tub-cd), that is a novel construct and
aresulted plasmid called pF4X1.4sat (tk- carries two "suicide" genes. The total size of our vector is 10 kb.
Conclusion: The two negative selection; HSV-tk and cd genes make the transformed cell sensitive to ganciclovir (GCV) and 5-fluorocytosine (5-FC). In future; we can use of theses recombinant Leishmania cells as the live recombinant vaccine strategy against Leishmaniasis.

 
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