Isolation of T-DNA flanking plant DNA from T-DNA insertional embryo-lethal mutants of Arabidopsis thaliana by plasmid rescue technique

کلن کردن ژنRGL2 از DNA ژنومی آرابیدوپسیس (Arabidopsis thaliana)

یکی از ژنهایی که در سنتز جیبرلین در گیاه شناخته شده و موقعیت آنها در ژنوم آرابیدپسیس مشخص شده است، RGL2 می‌باشد که در این بررسی کلن گردید. با استخراج DNA از آرابیدوپسیس، اکوتیپ کلمبیا و استفاده از آغازگرهای اختصاصی که بر اساس توالی ژن مورد نظر ساخته شده بودند، این ژن تکثیر شد. پلاسمید  PET-32a (+)و ژن تکثیر شده با استفاده از دو آنزیم برشی خاص به نامهای Nco I و Sal I برش داده شدند. پس از اتصال ژ...

FLAGdb/FST: a database of mapped flanking insertion sites (FSTs) of Arabidopsis thaliana T-DNA transformants

A large collection of T-DNA insertion transformants of Arabidopsis thaliana has been generated at the Institute of Agronomic Research, Versailles, France. The molecular characterisation of the insertion sites is currently performed by sequencing genomic regions flanking the inserted T-DNA (FST). The almost complete sequence of the nuclear genome of A.thaliana provides the framework for organisi...

GABI-Kat SimpleSearch: A Flanking Sequence Tag (FST) Database for the Identification of T-DNA Insertion Mutants in Arabidopsis Thaliana

SUMMARY GABI-Kat SimpleSearch is a database of flanking sequence tags (FSTs) of T-DNA mutagenized Arabidopsis thaliana lines that were generated by the GABI-Kat project. Sequences flanking the T-DNA insertion sites were aligned to the A.thaliana genome sequence, annotated with information about the FST, the insertion site and the line from which the FST was derived. A web interface permits text...

T-DNA as an insertional mutagen in Arabidopsis.

How effective is T-DNA insertional mutagenesis in Arabidopsis?

To evaluate the effectiveness of T-DNA insertion in knocking out a gene, 1084 published Arabidopsis thaliana insertion mutants representing 755 genes in 648 publications were reviewed. Insertion in the protein-coding region of a gene generates a knockout at least 90% of the time or 25% of the time if it is before the start codon which also produces a knockdown 67% of the time. Insertion after t...