inhibition of leishmania major ptr1 gene expression by anti-sense in escherichia coli
نویسندگان
چکیده
background: protozoa related to trypanosome family including leishmania , synthesize enzymes to escape from drug therapy. one of them is ptr1 that its enzymatic activity is similar to dihydrofolate reductase (dhfr). dihydrofolate reductase - thymidylate synthase has a major role in dna synthesis, if it is inhibited, the result would be the death of parasite. since ptr1 activity is similar to dhfr, causes the decrease of inhibition effect of drug. the aim of this study was inhibition of iranian l. major ptr1 expression with mrna antisense in prokaryotic system as an approach to appear of the drugs therapeutic effects more. methods: ptr1 gene was ligated to pacycduet-1 and pcdna3 plasmids as sense and antisense plasmids, respectively. simultaneously transfer of sense and antisense plasmids was done in e. coli strain m15. sds-page and western blot analysis were carried out to analyze the expression. results: sense and antisense plasmids were prepared and confirmed by restriction analysis and pcr then simultaneously transfer of them was done. sds-page and western blot analysis showed ptr1 gene was inhibited by mrna antisense in bacterial cells. conclusion: expression of ptr1 gene in sense plasmid was inhibited successfully by antisense plasmid.
منابع مشابه
Inhibition of Leishmania major PTR1 Gene Expression by Antisense in Escherichia coli
BACKGROUND Protozoa related to Trypanosome family including Leishmania, synthesize enzymes to escape from drug therapy. One of them is PTR1 that its enzymatic activity is similar to dihydrofolate reductase (DHFR). Dihydrofolate reductase - thymidylate synthase has a major role in DNA synthesis, if it is inhibited, the result would be the death of parasite. Since PTR1 activity is similar to DHFR...
متن کاملMolecular Cloning, Expression and Enzymatic Assay of Pteridine Reductase 1 from Iranian Lizard Leishmania
Background: Currently, there are no effective vaccines against leishmaniasis, and treatment using pentavalent antimonial drugs is occasionally effective and often toxic for patients. The PTR1 enzyme, which causes antifolate drug resistance in Leishmania parasites encoded by gene pteridine reductase 1 (ptr1). Since Leishmania lacks pteridine and folate metabolism, it cannot synthesize the pterid...
متن کاملSpecific Inhibition of the Expression of the Promyelocytic Leukemia (PML) Protein by Anti-Sense Oligonucleotides
In the present study, using anti-sense oligonucleotides the inhibition of expression of the PML protein hasbeen investigated. The anti-sense oligonucleotides were designed against the translation initiation site ofthe PML gene, and their effects were investigated on cellular growth and DNA synthesis. Incubation of normalhuman fibroblast cells with the anti-sense oligonucleotid...
متن کاملCrystallization of recombinant Leishmania major pteridine reductase 1 (PTR1).
The enzyme pteridine reductase (PTR1) has recently been discovered in the protozoan parasite Leishmania and validated as a target for therapeutic intervention. PTR1 is responsible for the salvage of pteridines and also contributes to antifolate drug resistance. Structural analysis, in combination with ongoing biochemical characterization will assist the elucidation of the structure-activity rel...
متن کاملCLONING AND EXPRESSION OF LEISHMANOLYSIN GENE FROM LEISHMANIA MAJOR IN PRIMATE CELL LINES
Leishmanolysin is a worldwide disease that is caused by different species of the genus Leishmania. Leishmanolysin, One of the genes expressed by Leishmania, appears to be an ideal candidate for genetic vaccination. In this study, a full length sequence, which encodes Leishmanolysin functionally critical regions (amino acids 100-579), was cloned from a Leishmania strain endemic to Iran. Analysis...
متن کاملExpression of Gp63 Gene from NIH Strain of Leishmania major in Pichia pastoris
Leishmaniasis is a major infectious disease of considerable public health in more than 86 countries around the world. Several approaches toward vaccine development against this disease have been taken. Glycoprotein (gp63) is conserved among diverse species of Leishmania and has induced immunological responses in murine models. Therefore, this glycoprotein has been considered as a second generat...
متن کاملمنابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
iranian journal of public healthجلد ۴۱، شماره ۶، صفحات ۶۵-۷۱
میزبانی شده توسط پلتفرم ابری doprax.com
copyright © 2015-2023