학술논문
Leishmania major ve Leishmania infantum Promastigot Formlarinin Karsilastirmali Gen Ekspresyon Profilleri/Comparative Gene Expression Profiles of Leishmania major and Leishmania infantum Promastigotes
Özgün Arastirma/Original Investigation
Özgün Arastirma/Original Investigation
Document Type
Academic Journal
Source
Turkish Journal of Parasitology. June 2021, Vol. 45 Issue 2, p88, 7 p.
Subject
Language
English
ISSN
1300-6320
Abstract
GIRIS Leishmaniasis, protozoa grubunda yer alan Leishmania parazitinin farkli türlerinin neden oldugu kompleks bir hastalik grubudur. Dünya Saglik Örgütü tarafindan yedi önemli tropikal hastaliktan biri olarak tanimlanan leishmaniasis, dünya genelinde [...]
Amaç: Bu çalismada, Leishmania major ve Leishmania infantum promastigotlarindaki gen ekspresyonlarinin karsilastirmali analizi yapilarak, iki tür arasinda gen ekspresyon profillerindeki farkliliklarin tespit edilmesi amaçlanmistir. Yöntemler: L. major (MHOM/IL/80) ve L. infantum (MHOM/MA/67/ITMAP/263) hücre hatlari kullanilarak hücre kültürü olusturulmustur. Daha sonra, total RNA izolasyonu ve cDNA sentezi gerçeklestirilerek; revers transkriptaz polimeraz zincir reaksiyonu ile metabolik yolaklarda ve nükleik asit sentezinde rol oynayan ve her iki türde ortak olan 30 genin ekspresyon seviyelerindeki kat degisimi hesaplanmistir. LeishDB ve KEGG veri tabanlari kullanilarak genlerin fonksiyonel islevleri belirlenmistir. Bulgular: Bu çalismada, L. major ile L. infantum promastigotlarinda ortak olarak eksprese edilen ve protein kodlayan 30 farkli gen profili degerlendirilmis ve iki tür arasinda anlamli farklilik saptanmistir (p1). Bu genler; phosphoglycan beta 1,3 galactosyltransferase-like, lathosterol oxidase-like, fatty acid elongase, 3-oxo-5 alpha-steroid 4-dehydrogenase, calpain-like cysteine peptidase, acetyl-coA synthetase, 3'-nucleotidase/nuclease, 3'-nucleotidase/nuclease precursor ve 3-ketoacyl-coA thiolase-like olarak belirlenmistir. Iki türde ortak olan genlerin karsilik geldigi proteinlerin fonksiyonlari veri tabanlarinda ayrintili olarak incelendiginde ise, bu genlerin parazitin lipit, protein ve karbonhidrat mekanizmalarinda, nükleik asit ve metabolizma fonksiyonlarinda rol oynadigi saptanmistir. Sonuç: L. major ile L. infantum türlerinde ortak olarak bulunan genlerin ekspresyon profillerindeki degisiklikler, parazit türleri arasinda virülans, patogenez, klinik ve tedavi farkliliklarina neden olabilir. Ayrica parazite karsi asi ve ilaç çalismalari için türlere özgü spesifik veya ortak hedeflerin seçilmesinde gen profillerinin degerlendirilmesi önem arz etmektedir. Anahtar Kelimeler: Gen ekpresyonu, kat degisimi, Leishmania infantum, Leishmania major, promastigote Objective: This study aimed to determine the differences between the gene expression profiles of Leismania major and Leishmania infantum promastigotes through comparative analysis of gene expressions. Methods: Cell culture of L. major (MHOM/IL/80) and L. infantum (MHOM/MA/67/ITMAP/263) cell lines was performed. Afterwards, total RNA isolation and cDNA synthesis were performed and fold changes in the expression levels of 30 genes that play a role in metabolic pathways and nucleic acid synthesis and co-expressed in two species were evaluated by reverse transcriptase polymerase chain reaction. Functions of genes were determined using LeishDB and KEGG databases. Results: In this study, profiles of protein-coding 30 genes expressed in L. major and L. infantum promastigotes were evaluated and significant differences were found between the two species (p1). These genes include phosphoglycan beta 1.3 galactosyltransferase-like, lathosterol oxidase-like, fatty acid elongase, 3-oxo-5 alpha-steroid 4-dehydrogenase, calpain-like cysteine peptidase, acetyl-coA synthetase, 3'-nucleotidase/nuclease, 3'-nucleotidase/nuclease precursor and 3-ketoacyl-coA thiolase-like. When the functions of the proteins that correspond to the genes common in the two species were examined in detail using the databases, it was determined that these genes play role in lipid, protein, carbohydrate and nucleic acid metabolic functions of the parasite. Conclusion: Alterations in the expression profiles of genes common to L. major and L. infantum species may cause differences in the virulence, pathogenesis, clinical features and treatment modality between these parasite species. In addition, evaluation of gene profiles is important in the selection of speciesspecific or common targets for vaccine and drug studies. Keywords: Gene expression, fold change, Leishmania infantum, Leishmania major, promastigot
Amaç: Bu çalismada, Leishmania major ve Leishmania infantum promastigotlarindaki gen ekspresyonlarinin karsilastirmali analizi yapilarak, iki tür arasinda gen ekspresyon profillerindeki farkliliklarin tespit edilmesi amaçlanmistir. Yöntemler: L. major (MHOM/IL/80) ve L. infantum (MHOM/MA/67/ITMAP/263) hücre hatlari kullanilarak hücre kültürü olusturulmustur. Daha sonra, total RNA izolasyonu ve cDNA sentezi gerçeklestirilerek; revers transkriptaz polimeraz zincir reaksiyonu ile metabolik yolaklarda ve nükleik asit sentezinde rol oynayan ve her iki türde ortak olan 30 genin ekspresyon seviyelerindeki kat degisimi hesaplanmistir. LeishDB ve KEGG veri tabanlari kullanilarak genlerin fonksiyonel islevleri belirlenmistir. Bulgular: Bu çalismada, L. major ile L. infantum promastigotlarinda ortak olarak eksprese edilen ve protein kodlayan 30 farkli gen profili degerlendirilmis ve iki tür arasinda anlamli farklilik saptanmistir (p1). Bu genler; phosphoglycan beta 1,3 galactosyltransferase-like, lathosterol oxidase-like, fatty acid elongase, 3-oxo-5 alpha-steroid 4-dehydrogenase, calpain-like cysteine peptidase, acetyl-coA synthetase, 3'-nucleotidase/nuclease, 3'-nucleotidase/nuclease precursor ve 3-ketoacyl-coA thiolase-like olarak belirlenmistir. Iki türde ortak olan genlerin karsilik geldigi proteinlerin fonksiyonlari veri tabanlarinda ayrintili olarak incelendiginde ise, bu genlerin parazitin lipit, protein ve karbonhidrat mekanizmalarinda, nükleik asit ve metabolizma fonksiyonlarinda rol oynadigi saptanmistir. Sonuç: L. major ile L. infantum türlerinde ortak olarak bulunan genlerin ekspresyon profillerindeki degisiklikler, parazit türleri arasinda virülans, patogenez, klinik ve tedavi farkliliklarina neden olabilir. Ayrica parazite karsi asi ve ilaç çalismalari için türlere özgü spesifik veya ortak hedeflerin seçilmesinde gen profillerinin degerlendirilmesi önem arz etmektedir. Anahtar Kelimeler: Gen ekpresyonu, kat degisimi, Leishmania infantum, Leishmania major, promastigote Objective: This study aimed to determine the differences between the gene expression profiles of Leismania major and Leishmania infantum promastigotes through comparative analysis of gene expressions. Methods: Cell culture of L. major (MHOM/IL/80) and L. infantum (MHOM/MA/67/ITMAP/263) cell lines was performed. Afterwards, total RNA isolation and cDNA synthesis were performed and fold changes in the expression levels of 30 genes that play a role in metabolic pathways and nucleic acid synthesis and co-expressed in two species were evaluated by reverse transcriptase polymerase chain reaction. Functions of genes were determined using LeishDB and KEGG databases. Results: In this study, profiles of protein-coding 30 genes expressed in L. major and L. infantum promastigotes were evaluated and significant differences were found between the two species (p1). These genes include phosphoglycan beta 1.3 galactosyltransferase-like, lathosterol oxidase-like, fatty acid elongase, 3-oxo-5 alpha-steroid 4-dehydrogenase, calpain-like cysteine peptidase, acetyl-coA synthetase, 3'-nucleotidase/nuclease, 3'-nucleotidase/nuclease precursor and 3-ketoacyl-coA thiolase-like. When the functions of the proteins that correspond to the genes common in the two species were examined in detail using the databases, it was determined that these genes play role in lipid, protein, carbohydrate and nucleic acid metabolic functions of the parasite. Conclusion: Alterations in the expression profiles of genes common to L. major and L. infantum species may cause differences in the virulence, pathogenesis, clinical features and treatment modality between these parasite species. In addition, evaluation of gene profiles is important in the selection of speciesspecific or common targets for vaccine and drug studies. Keywords: Gene expression, fold change, Leishmania infantum, Leishmania major, promastigot