Screening for glutamine synthetase knockout mutants in Arabidopsis
2011
Аутори:
Dragićević, MilanBogdanović, Milica
Todorović, Slađana
Ghalawenji, Nabil
Simonović, Ana
Grubišić, Dragoljub
Остала ауторства
Vinterhalter, DraganТип документа:
Конференцијски прилог (Објављена верзија)
,
© 2011 by the Serbian Plant Physiology Society
Метаподаци
Приказ свих података о документуАпстракт:
Glutamine synthetase (GS, EC 6.3.1.2) is a key enzyme of ammonium assimilation and recycling in plants.
In Arabidopsis, five GLN1 genes encode cytosolic (GS1) isoforms which combine to form, at least nine multimeric GS holoenzymes with overlapping expression. To investigate the role of each isoform, we used respective T-DNA insertion (SALK and SAIL) lines, obtained from The European Arabidopsis Stock Centre. The lines were picked so that the insertion is located in the exons of GLN genes, hopefully preventing the functionality of the transcripts. The plants carrying the T-DNA insertion were selected on kanamycin (for SALK lines carrying the neo gene) or phosphinotricin (for SAIL lines carrying the bar gene). The surviving plants were screened with a three-primer PCR protocol which allows for differentiation of wild type, homozygous and heterozygous plants. For monitoring the transcription of wild type and mutant alleles, the fast, simple and cost-efficient Touch and Go technique was used. So far, homozygous knockout mutants of two high expressed GS isoforms (GLN1;2 and GLN1;3) have been identified.
Кључне речи:
Glutamine synthetase; ammonium assimilation; Arabidopsis; SALK and SAIL linesФинансирање / пројекти:
- Физиолошка, хемијска и молекуларна анализа диверзитета одабраних ретких и угрожених биљних врста у циљу еx ситу заштите и продукције биолошки активних једињења (RS-MESTD-Basic Research (BR or ON)-173024)
У:
- Vinterhalter D, editor. Programme and Abstracts: 19th Symposium of the Serbian Plant Physiology Society; 2011 Jun 13-15; Banja Vrujci, Serbia. Belgrade: Serbian Plant Physiology Society; 2022. p. 49.