کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5916113 | 1163366 | 2007 | 13 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Construction and analysis of Leishmania tarentolae transgenic strains free of selection markers
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
ODCRFUadenine phosphoribosyltransferaseNourseothricinNLSLeishmania tarentolaeHYGNEOSSUTETAPRTSATeGFPTMPNTCRNA polymerase II - آرانای پلیمراز II T7 RNA polymerase - T7 RNA پلیمرازBle - بودtetracycline repressor - تتراسایکلین سرکوبگرTetracycline - تتراسایکلینthymidine monophosphate - تیمیدین مونوفسفاتthymidine kinase - تیمیدین کینازnuclear localization signal - سیگنال محلی سازی هسته ایAntibiotic resistance - مقاومت به آنتی بیوتیکUTR یا untranslated regions - منطقه ترجمه نشدهuntranslated region - منطقه غیر ترجمهIntergenic region - منطقه مصنوعیHomologous recombination - نوترکیبی همولوگrelative fluorescent units - واحدهای فلورسنت نسبیtub - وانPol II - پل دومβ-Tubulin gene - ژن β-توبولینNeomycin resistance gene - ژن مقاومت نئومایسینhygromycin resistance gene - ژن مقاومت هیگرموسین
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شناسی مولکولی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Construction and analysis of Leishmania tarentolae transgenic strains free of selection markers Construction and analysis of Leishmania tarentolae transgenic strains free of selection markers](/preview/png/5916113.png)
چکیده انگلیسی
The trypanosomatid protozoan Leishmania tarentolae has been extensively used as a model system for studying causative agents of several tropical diseases and more recently as a host for recombinant protein production. Here we analyze the rates of partial or complete deletions of expression cassettes integrated into small ribosomal RNA and tubulin gene clusters as well as into ornithine decarboxylase gene of L. tarentolae. In approximately 60% of cases gene conversion was responsible for the deletion while in the rest of the cases deletion occurred within the expression cassette. We used this observation to design constitutive and inducible expression vectors that could be stably integrated into the genome and subsequently depleted of the antibiotic resistance genes using thymidine kinase or bleomycin resistance genes as negative selection markers. This enabled us to obtain L. tarentolae strains containing constitutive or inducible markerless expression cassettes. Analysis of the markerless strains demonstrated that although stability varied among clones some were stable for as many as 200 generations. We expect that this approach will be useful for the construction of strains carrying multiple expression cassettes for analysis of trypanosomatid pathogenicity mechanisms and overexpression of multi-subunit protein complexes for biochemical and structural studies.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Molecular and Biochemical Parasitology - Volume 155, Issue 2, October 2007, Pages 71-83
Journal: Molecular and Biochemical Parasitology - Volume 155, Issue 2, October 2007, Pages 71-83
نویسندگان
Sergei Mureev, Susanna Kushnir, Alexander A. Kolesnikov, Reinhard Breitling, Kirill Alexandrov,