کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2010027 1066706 2008 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Self-EcoTILLING to identify single-nucleotide mutations in multigene family
موضوعات مرتبط
علوم زیستی و بیوفناوری علوم کشاورزی و بیولوژیک علوم زراعت و اصلاح نباتات
پیش نمایش صفحه اول مقاله
Self-EcoTILLING to identify single-nucleotide mutations in multigene family
چکیده انگلیسی

TILLING (Targeting Induced Local Lesions IN Genomes) is a low-cost, high-throughput reverse genetic technique that employs a mismatch-specific endonuclease CEL-1 to discover induced point mutations in the genes of interest. The use of the TILLING technique to survey natural variation in genes is called EcoTILLING. Here, we report a modified EcoTILLING method for the discovery of mutations in multigene family, which we coin “Self-EcoTILLING” by using an allotetraploid Monochoria vaginalis ALS multigene family as an example. The mutations could be detected by TILLING of PCR products resulting from the primers specific to both Als1 and Als3 without involving the experimental step of mixture of reference and query DNA. Either of the two co-amplified loci could serve as reference DNA to the other. We demonstrate with this example that Self-EcoTILLING is a fast, reliable and economical technique of detecting single-nucleotide mutations in polyploid plants containing multigene family.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Pesticide Biochemistry and Physiology - Volume 92, Issue 1, September 2008, Pages 24–29
نویسندگان
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