کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5591055 | 1570334 | 2017 | 29 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
An efficient system for the generation of marked genetic mutants in members of the genus Burkholderia
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کلمات کلیدی
CMRTCrMCstrimethoprim resistanceBHRCMSKmr - KMRSuicide vector - بردار خودکشیTIR - تیرTPR - روش پاسخ فیزیکیmultiple cloning site - سایت کلونینگ چندگانهGene inactivation - غیر فعال سازی ژنBurkholderia cepacia complex - مجتمع Burkholderia cepaciaAntibiotic-resistance - مقاومت آنتی بیوتیکیTetracycline resistance - مقاومت تتراسیکلینKanamycin resistance - مقاومت کانامایسینTranslation initiation region - منطقه شروع ترجمه
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
To elucidate the function of a gene in bacteria it is vital that targeted gene inactivation (allelic replacement) can be achieved. Allelic replacement is often carried out by disruption of the gene of interest by insertion of an antibiotic-resistance marker followed by subsequent transfer of the mutant allele to the genome of the host organism in place of the wild-type gene. However, due to their intrinsic resistance to many antibiotics only selected antibiotic-resistance markers can be used in members of the genus Burkholderia, including the Burkholderia cepacia complex (Bcc). Here we describe the construction of improved antibiotic-resistance cassettes that specify resistance to kanamycin, chloramphenicol or trimethoprim effectively in the Bcc and related species. These were then used in combination with and/or to construct a series enhanced suicide vectors, pSHAFT2, pSHAFT3 and pSHAFT-GFP to facilitate effective allelic replacement in the Bcc. Validation of these improved suicide vectors was demonstrated by the genetic inactivation of selected genes in the Bcc species Burkholderia cenocepacia and B. lata, and in the non-Bcc species, B. thailandensis.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Plasmid - Volume 89, January 2017, Pages 49-56
Journal: Plasmid - Volume 89, January 2017, Pages 49-56
نویسندگان
Sravanthi Shastri, Helena L. Spiewak, Aderonke Sofoluwe, Vigdis A. Eidsvaag, Atif H. Asghar, Tyrone Pereira, Edward H. Bull, Aaron T. Butt, Mark S. Thomas,