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Caracteriza????o funcional do promotor de soja UCES8.3Lins, Philippe de Castro 10 April 2015 (has links)
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Previous issue date: 2015-04-10 / Gene promoters regulate gene expression quantitatively and qualitatively. The
regulatory sequences have cis elements and with trans acting that will direct and
correctly position the RNA polymerase which is the process of DNA transcription.
Genetic engineering of plants by transforming plants expressing genes of interest,
use in most studies, constitutive CaMV35S promoter character. A new promoter
isolated from soybeans, which is called uceS8.3 showing a constitutive promoter
driving expression in different plant tissues. The analysis of the expression of the
uidA gene, GUS, demonstrated that the expression profile uceS8.3 controlled by the
promoter is comparable or superior to the CaMV35S promoter in plant tissues such
as flower buds and roots. Were identified following the uceS8.3 promoter cis
regulatory elements that may be responsible for gene expression profile controlled by
this promoter. Modules of this promoter, when compared with the CaMV35S promoter
and the uceS8.3 itself demonstrated a difference in expression in plant tissues. Cis
regulatory elements as ROOTMOTIFPABOX1, POLLEN1LELAT52, MRE, Sp1 and Ibox.
The set of specific cis elements located in the promoter uceS8.3 modules may
be the minimum necessary elements to control expression in leaf and flower bud
tissues. The quantitative expression analysis and fluorometric GUS assays leaf and
root tissues have demonstrated a correlation between transcript levels and the
specific activity levels of a ??-glucuronidase enzyme in module 2 (720pb), but there is
a difference in the balance of these levels between uceS8.3 promoter and Module 4
(170pb). The studies presented here demonstrated that the promoter and its modules
has high ability to drive expression in flower tissues and roots, may be used and
applied to different types of biotechnologically biotic stresses, including insect pests
and nematodes. / Promotores g??nicos regulam a express??o de genes, quantitativamente e
qualitativamente. As sequ??ncias regulat??rias possuem elementos cis e trans
atuantes que v??o direcionar e posicionar corretamente a RNA polimerase para que
haja o processo de transcri????o do DNA. A engenharia gen??tica de plantas, por meio
da transforma????o de plantas, expressando genes de interesse, vem utilizando, na
maioria dos estudos, o promotor CaMV35S de car??ter constitutivo. Um novo
promotor isolado de soja, denominado uceS8.3 ?? tamb??m um promotor constitutivo
demonstrando conduzir a express??o em diferentes tecidos vegetais. A an??lise da
express??o do gene uidA, GUS, demonstrou que o perfil de express??o controlada
pelo promotor uceS8.3 ?? compar??vel ou superior ao promotor CaMV35S em tecidos
vegetais, como bot??o floral e raiz. Foram identificados, na sequ??ncia do promotor
uceS8.3, elementos cis regulat??rios que podem ser respons??veis pelo perfil de
express??o g??nica controlada por esse promotor. Os m??dulos desse promotor,
quando comparados com o promotor CaMV35S, e o pr??prio uceS8.3 demonstraram
uma diferen??a de express??o nos tecidos vegetais. Elementos cis regulat??rios como
ROOTMOTIFPABOX1, POLLEN1LELAT52, MRE, Sp1 e I-box. O conjunto de
elementos cis espec??ficos, localizados nos m??dulos do promotor uceS8.3, podem ser
os elementos m??nimos necess??rios para controlar a express??o em tecidos de folha e
bot??o floral. A an??lise de express??o quantitativa e de ensaios fluorim??tricos de GUS
nos tecidos folha e raiz, demonstraram uma correla????o entre os n??veis de transcrito
e os n??veis de atividade espec??fica da enzima ??-glucuronidase no m??dulo 2 (720pb),
por??m h?? uma diferen??a na correla????o destes n??veis entre o promotor uceS8.3 e o
m??dulo 4 (170pb). Os estudos aqui apresentados demonstraram que o promotor
uceS8.3 e seus m??dulos possuem alta capacidade de conduzir express??o em
tecidos florais e ra??zes, podendo ser utilizado e aplicado biotecnologicamente para
diferentes tipos de estresses bi??ticos, incluindo insetos-praga e nemat??ides.
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