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Pesquisa : G05.728.865 [Categoria DeCS]
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Id: biblio-1053457
Autor: Suwanseree, Valerie; Phansiri, Salak; Yapwattanaphun, Chinawat.
Título: A comparison of callus induction in 4 Garcinia species
Fonte: Electron. j. biotechnol;40:45-51, July. 2019. ilus, tab.
Idioma: en.
Resumo: Background: This research is intended to determine suitable types and concentrations of plant growth regulators (PGRs) to induce callus on stem and leaf sections of 4 species of the genus Garcinia, namely, Garcinia mangostana, Garcinia schomburgkiana, Garcinia cowa, and Garcinia celebica. The base medium was MS medium containing 30 g l -1 sucrose, 0.5 g l-1 polyvinylpyrrolidone (PVP), and 7 g l-1 agar, and for the different treatments, PGRs were added to the medium as follows: thidiazuron (TDZ) at concentrations of 0, 0.1, 0.5, 1, and 2 mg l-1; 6-(3- hydroxybenzylamino) purine (meta-topolin) at concentrations of 0, 0.5, 2.5, and 5 mg l-1; 4-amino-3,5,6- trichloro-2-pyridinecarboxylic acid (picloram) at concentrations of 0, 0.5, 2.5, and 5 mg l-1; and 2,4- dichlorophenoxyacetic acid (2,4-D) at concentrations of 0, 0.5, 1, 2, and 4 mg l-1. The occurrence of callus was observed after 4 weeks. Results: A maximum of 100% and 93% of G. mangostana leaf explants formed callus in the 0.5 mg l-1 and 1 mg l-1 TDZ treatments, respectively, while 100% of G. schomburgkiana stem explants formed callus in the 1 mg l-1 TDZ treatment and 89% of G. schomburgkiana leaf explants formed callus in the 0.5 mg l-1 picloram treatment. The highest callus induction rate for G. cowa was 62% in the 1 mg l-1 TDZ treatment and for G. celebica was 56% in the 0.5 mg l-1•mT-1 treatment. Conclusions: For all 4 species, the greatest amount of large nodular callus was observed in the TDZ treatments. White, friable callus was observed on most of the 2,4-D and picloram treatment groups. Most meta-topolin treatments resulted in minimal callus formation.
Descritores: Reguladores de Crescimento de Planta/metabolismo
Garcinia/crescimento & desenvolvimento
Compostos Fitoquímicos/metabolismo
-Compostos de Fenilureia
Tiadiazóis
Fatores de Tempo
Transformação Genética
Clusiaceae/crescimento & desenvolvimento
Garcinia/fisiologia
Técnicas de Cultura de Tecidos
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1087255
Autor: Arencibia, Ariel D; D'Afonseca, Vívian; Chakravarthi, Mohan; Castiglione, Stefano.
Título: Learning from transgenics: advanced gene editing technologies should also bridge the gap with traditional genetic selection
Fonte: Electron. j. biotechnol;41:22-29, sept. 2019. ilus.
Idioma: en.
Resumo: We highlight the importance of the mixed genetic approaches (classical and currents) to improve the social perception related to the GMOs acceptance. We pointed out that CRISPR/Cas9 events could carry DNA variability/rearrangements related to somaclonal variations or epigenetic changes that are independent from the editing per se. The transformation of single cells, followed by plant regeneration, is used to generate modified plants, transgenic or genome editing (CRISPR/Cas9). The incidence of undesirable somaclonal variations and/or epigenetic changes that might have occurred during in vitro multiplication and regeneration processes, must be carefully analyzed in replicates in field trials. One remarkable challenge is related to the time lapse that selects the modified elite genotypes, because these strategies may spend a variable amount of time before the results are commercialized, where in all the cases it should be take into account the genotype × environment interactions. Furthermore, this combination of techniques can create an encouraging bridge between the public opinion and the community of geneticists who are concerned with plant genetic improvement. In this context, either transgenesis or genomic editing strategies become complementary modern tools to facing the challenges of plant genetic improvement. Their applications will depend on case-by-case analysis, and when possible will necessary associate them to the schemes and bases of classic plant genetic improvement.
Descritores: Plantas Geneticamente Modificadas
Técnicas de Transferência de Genes
Sistemas CRISPR-Cas
Edição de Genes
-Transformação Genética
Mutagênese
Metilação de DNA
Melhoramento Genético
Epigênese Genética
Tipo de Publ: Revisão
Responsável: CL1.1 - Biblioteca Central


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Id: lil-129321
Autor: Villa, Luisa.
Título: Epidemiología molecular y evolución de los papiloma virus humanos / Human Papillomavirus molecular epidemiology and evolution
Fonte: Acta cancerol;23(3):13-8, set. 1993. ilus, tab, mapas.
Idioma: es.
Resumo: Hemos amplificado, clonado y secuenciado por la reacción en cadena de la polimerasa, los segmentos genómicos de 118 papiloma virus humanos tipo 16 (PVH-16) aislados de 76 biopsias cervicales, 14 exstendidos cervicales, 3 biopsias vulvares, 2 biopsias de pene, 2 biopsias anales, 1 biopsia vaginal y dos líneas celulares. Los especímenes fueron obtenidos de pacientes en cuatro países- Singapur, Brasil, Tanzania y Alemania. La secuencia de un fragmento de 364-bp de la región de control larga (RLC) de virus reveló métodos de distancia matriz y el enfoque de series de transformación. Los árboles basados en la RLV fueron confirmados por otro sset basado en la región completa. E5. ambos sets tuvieron dos ramas. Casi todas las variantes de Tanzania fueron asignadas a la rama africana, y todas las alemanas y al mayoría de las variantes de Singapur lo fueron a la rama eurasiática. En contraste con la homogeneidad interna de las variantes de Singapur Alemania y Tanzania, las variantes de Brasil estuvieron claramente divididas en las dos ramas. Los datos sugieren que el PVH-16 evolucionó separadamente por un tiempo largo en Africa y Eurasia. Representantes de ambas ramas podrían haber sido transferidos a Brasil vía la inmigración colonial. Representantes de la rama africana fueron posiblemente llevados al Lejano Este a través de las viejas rutas marinas árabes e indonesias. Nuestro estudio da soportes a la idea de que el PVH-16 es un tipo bien definido, pues las variantes muestran una divergencia genómica máxima de alrededor de 5 por ciento. La pequeña divergencia en cada localización geográfica y la falta de divergencia marcada entre las variantes genómica de Tanzania y la Brasilera-Africana luego de doscientos años desde su posible introducción en el Nuevo Mundo, sugieren una tasa muy lenta de evolución viral. El árbol filogenético probablemente representa un mínimo de varias centurias de evolución, si no una de igual a la de la raza humana.
Descritores: Papillomaviridae/genética
Variação Genética
Evolução Biológica
-Papillomaviridae/isolamento & purificação
Transformação Genética
Limites: Humanos
Responsável: PE1.1 - Oficina Universitária de Biblioteca


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Id: biblio-1022493
Autor: Liu, Dongmei; Zhu, Hanyu; Chen, Yue; Zheng, Liesheng; Chen, Liguo; Ma, Aimin.
Título: Cloning and heterologous expression of a hydrophobin gene Ltr. hyd from the tiger milk mushroom Lentinus tuber-regium in yeast-like cells of Tremella fuciformis
Fonte: Electron. j. biotechnol;32:6-12, Mar. 2018. tab, graf, ilus.
Idioma: en.
Projeto: National Natural Science Foundation of China (NSFC).
Resumo: Background: Hydrophobins are small proteins secreted by filamentous fungi, which show a highly surface activity. Because of the signally self-assembling abilities and surface activities, hydrophobins were considered as candidates in many aspects, for example, stabilizing foams and emulsions in food products. Lentinus tuber-regium, known as tiger milk mushroom, is both an edible and medicinal sclerotium-producing mushroom. Up to now, the hydrophobins of L. tuber-regium have not been identified. Results: In this paper, a Class I hydrophobin gene, Ltr.hyd, was cloned from L. tuber-regium and expressed in the yeast-like cells of Tremella fuciformis mediated by Agrobacterium tumefaciens. The expression vector pGEH-GH was under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter. The integration of Ltr.hyd into the genome of T. fuciformis was confirmed by PCR, Southern blot, fluorescence observation and quantitative real-time PCR (qRT-PCR). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that recombinant hydrophobin rLtr.HYD with an expected molecular mass of 13 kDa was extracted. The yield of rLtr.HYD was 0.66 mg/g dry weight. The emulsifying activity of rLtr.HYD was better than the typical food emulsifiers sodium caseinate and Tween 20. Conclusions: We evaluated the emulsifying property of hydrophobin Ltr.HYD, which can be potentially used as a food emulsifier.
Descritores: Basidiomycota/metabolismo
Proteínas Fúngicas/genética
Lentinula/genética
Lentinula/metabolismo
-Transformação Genética
Basidiomycota/enzimologia
Leveduras
Proteínas Fúngicas/metabolismo
Southern Blotting
Clonagem Molecular
Agrobacterium tumefaciens/metabolismo
Análise de Sequência
Emulsificantes
Eletroforese em Gel de Poliacrilamida
Reação em Cadeia da Polimerase em Tempo Real
Gliceraldeído-3-Fosfato Desidrogenases/metabolismo
Microscopia de Fluorescência
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1021034
Autor: Mihálik, Daniel; Gubisová, Marcela; Kraic, Ján; Hudcovicová, Martina; Havrlentová, Michaela; Moravcíková, Jana; Glasa, Miroslav; Matusíková, Ildikó.
Título: Introduction of a synthetic Thermococcus-derived α-amlyase gene into barley genome for increased enzyme thermostability in grains
Fonte: Electron. j. biotechnol;30:1-5, nov. 2017. ilus, tab, graf.
Idioma: en.
Projeto: Slovak Research and Development Agency.
Resumo: Background: The enzymes utilized in the process of beer production are generally sensitive to higher temperatures. About 60% of them are deactivated in drying the malt that limits the utilization of starting material in the fermentation process. Gene transfer from thermophilic bacteria is a promising tool for producing barley grains harboring thermotolerant enzymes. Results: Gene for α-amylase from hydrothermal Thermococcus, optimally active at 75­85°C and pH between 5.0 and 5.5, was adapted in silico to barley codon usage. The corresponding sequence was put under control of the endosperm-specific promoter 1Dx5 and after synthesis and cloning transferred into barley by biolistics. In addition to model cultivar Golden Promise we transformed three Slovak barley cultivars Pribina, Levan and Nitran, and transgenic plants were obtained. Expression of the ~50 kDa active recombinant enzyme in grains of cvs. Pribina and Nitran resulted in retaining up to 9.39% of enzyme activity upon heating to 75°C, which is more than 4 times higher compared to non-transgenic controls. In the model cv. Golden Promise the grain α-amylase activity upon heating was above 9% either, however, the effects of the introduced enzyme were less pronounced (only 1.22 fold difference compared with non-transgenic barley). Conclusions: Expression of the synthetic gene in barley enhanced the residual α-amylase activity in grains at high temperatures.
Descritores: Sementes/enzimologia
Hordeum/enzimologia
Thermococcus/metabolismo
alfa-Amilases/metabolismo
-Sementes/genética
Sementes/microbiologia
Transformação Genética
Hordeum/genética
Hordeum/microbiologia
Cerveja
Estabilidade Enzimática
Plantas Geneticamente Modificadas/enzimologia
Clonagem Molecular
Técnicas de Transferência de Genes
alfa-Amilases/genética
Fermentação
Termotolerância
Temperatura Alta
Concentração de Íons de Hidrogênio
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1017249
Autor: Li, Tao; Ding, Yatong; Zhang, Jun; Jiao, Guobao; Sun, Lipeng; Liu, Zhongmin; Qiu, Liyou.
Título: Improving the expression of recombinant pullulanase by increasing mRNA stability in Escherichia coli
Fonte: Electron. j. biotechnol;29:63-67, sept. 2017. ilus, tab, graf.
Idioma: en.
Projeto: National Science and Technology Program in Rural Areas during the 12th Five-Year Plan period.
Resumo: Background: Pullulanase production in both wild-type strains and recombinantly engineered strains remains low. The Shine-Dalgarno (SD) sequence and stem-loop structure in the 5' or 3' untranslated region (UTR) are well-known determinants of mRNA stability. This study investigated the effect of mRNA stability on pullulanase heterologous expression. Results: We constructed four DNA fragments, pulA, SD-pulA, pulA-3t, and SD-pulA-3t, which were cloned into the expression vector pHT43 to generate four pullulanase expression plasmids. The DNA fragment pulA was the coding sequence (CDS) of pulA in Klebsiella variicola Z-13. SD-pulA was constructed by the addition of the 5' SD sequence at the 5' UTR of pulA. pulA-3t was constructed by the addition of a 3' stem-loop structure at the 3' UTR of pulA. SD-pulA-3t was constructed by the addition of the 5' SD sequence at the 5' UTR and a 3' stem-loop structure at the 3' UTR of pulA. The four vectors were transformed into Escherichia coli BL21(DE3). The pulA mRNA transcription of the transformant harboring pHT43-SD-pulA-3t was 338.6%, 34.9%, and 79.9% higher than that of the other three transformants, whereas the fermentation enzyme activities in culture broth and intracellularly were 107.0 and 584.1 times, 1.2 and 2.0 times, and 62.0 and 531.5 times the amount of the other three transformants (pulA, SD-pulA, and pulA-3 t), respectively. Conclusion: The addition of the 5' SD sequence at the 5' UTR and a 3' stem-loop structure at the 3' UTR of the pulA gene is an effective approach to increase pulA gene expression and fermentation enzyme activity.
Descritores: Escherichia coli/enzimologia
Escherichia coli/genética
Glicosídeo Hidrolases/metabolismo
-Transformação Genética
Expressão Gênica
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Estabilidade de RNA
Fermentação
Vetores Genéticos
Glicosídeo Hidrolases/genética
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1009000
Autor: Liu, Chunming; Yang, Bowen; Ming, Yuetong; Liu, Jianfeng; Cheng, Yunqing.
Título: Construction of an RNAi vector for knockdown of GM-ACS genes in the cotyledonary nodes of soybean
Fonte: Electron. j. biotechnol;26:40-45, Mar. 2017. ilus, graf.
Idioma: en.
Projeto: National Natural Science Foundation of China; . Special Foundation for Young Scientists of Jilin Province, China.
Resumo: Background: Ethylene plays an important role in the regulation of floral organ development in soybean, and 1-aminocyclopropane-1-carboxylate synthase (ACS) is a rate-limiting enzyme for ethylene biosynthesis. However, whether ACS also regulates floral organ differentiation in soybean remains unknown. To address this, we constructed an RNAi vector to inhibit ACS expression in cotyledonary nodes. Linear DNA cassettes of RNAi-ACS obtained by PCR were used to transform soybean cotyledonary nodes. Results: In total, 131 of 139 transiently transformed plants acquired herbicide resistance and displayed GUS activities in the new buds. In comparison to untransformed seedling controls, a greater number of flower buds were differentiated at the cotyledonary node; GM-ACS1 mRNA expression levels and ethylene emission in the transformed buds were reduced. Conclusion: These results indicate that the cotyledonary node transient transformation system may be suitable for stable transformation and that the inhibition of ACS expression may be an effective strategy for promoting floral organ differentiation in soybean.
Descritores: Soja/enzimologia
Soja/genética
Interferência de RNA
Liases/metabolismo
-Soja/crescimento & desenvolvimento
Transformação Genética
Expressão Gênica
Diferenciação Celular
Reação em Cadeia da Polimerase
Regulação da Expressão Gênica de Plantas
Etilenos/biossíntese
Resistência a Herbicidas
Vetores Genéticos
Glucuronidase
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1008414
Autor: Liu, Juhua; Gao, Pengzhao; Sun, Xiuxiu; Zhang, Jing; Sun, Peiguang; Wang, Jiashui; Jia, Caihong; Zhang, Jianbin; Hu, Wei; Xu, Biyu; Jin, Zhiqiang.
Título: Efficient regeneration and genetic transformation platform applicable to five Musa varieties
Fonte: Electron. j. biotechnol;25:33-38, ene. 2017. tab, ilus.
Idioma: en.
Projeto: Modern Agro-industry Technology Research System; . National Nonprofit Institute Research Grant of Institute of Tropical Bioscience and Biotechnology CATAS-ITBB.
Resumo: Background: Banana (Musa spp.) is an important staple food, economic crop, and nutritional fruit worldwide. Conventional breeding has been seriously hampered by their long generation time, polyploidy, and sterility of most cultivated varieties. Establishment of an efficient regeneration and transformation system for banana is critical to its genetic improvement and functional genomics. Results: In this study, a vigorous and repeatable transformation system for banana using direct organogenesis was developed. The greatest number of shoots per explant for all five Musa varieties was obtained using Murashige and Skoog medium supplemented with 8.9 µM benzylaminopurine and 9.1 µM thidiazuron. One immature male flower could regenerate 380­456, 310­372, 200­240, 130­156, and 100­130 well-developed shoots in only 240­270 d for Gongjiao, Red banana, Rose banana, Baxi, and Xinglongnaijiao, respectively. Longitudinal sections of buds were transformed through particle bombardment combined with Agrobacterium-mediated transformation using a promoterless ß-glucuronidase (GUS) reporter gene; the highest transformation efficiency was 9.81% in regenerated Gongjiao plantlets in an optimized selection medium. Transgenic plants were confirmed by a histochemical assay of GUS, polymerase chain reaction, and Southern blot. Conclusions: Our robust transformation platform successfully generated hundreds of transgenic plants. Such a platform will facilitate molecular breeding and functional genomics of banana.
Descritores: Musa/crescimento & desenvolvimento
Musa/genética
-Regeneração
Transformação Genética
Imuno-Histoquímica
Southern Blotting
Reação em Cadeia da Polimerase
Plantas Geneticamente Modificadas
Agrobacterium tumefaciens/fisiologia
Musa/microbiologia
Organogênese Vegetal
Glucuronidase
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1017075
Autor: Yu, Xiao-Chun; Ma, Shi-Liang; Xu, Yan; Fu, Cheng-Hao; Jiang, Chun-Ying; Zhou, Chen-Yu.
Título: Construction and application of a novel genetically engineered Aspergillus oryzae for expressing proteases
Fonte: Electron. j. biotechnol;29:32-38, sept. 2017. tab, ilus, graf.
Idioma: en.
Resumo: Background: We aimed to test the possibility of improving polypeptide production from soybean meal fermentation by engineered Aspergillus oryzae strains. Four different protease genes were cloned and transformed into wild-type A. oryzae, and the engineered A. oryzae strains were then used for soybean meal fermentation. Results: The results showed different degrees of improvement in the protease activity of the four transformants when compared with wild-type A. oryzae. A major improvement in the polypeptide yield was achieved when these strains were used in soybean meal fermentation. The polypeptide conversion rate of one of the four transformants, A. oryzae pep, reached 35.9%, which was approximately twofold higher than that exhibited by wild-type A. oryzae. Amino acid content analysis showed that the essential amino acid content and amino acid composition of the fermentation product significantly improved when engineered A. oryzae strains were used for soybean meal fermentation. Conclusions: These findings suggest that cloning of microbial protease genes with good physicochemical properties and expressing them in an ideal host such as A. oryzae is a novel strategy to enhance the value of soybean meal.
Descritores: Peptídeo Hidrolases/metabolismo
Aspergillus oryzae/enzimologia
Aspergillus oryzae/genética
-Peptídeo Hidrolases/genética
Soja
Transformação Genética
Engenharia Genética
Clonagem Molecular
Fermentação
Farinha
Aminoácidos/análise
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-914354
Autor: Silva, André Luís Lopes da; Oliveira, Yohana de; Procopiuk, Marcia; Mudry, Clarissa de Souza; Brondani, Gilvano Ebling; Costa, Jefferson da Luz; Scheidt, Gessiel Newton.
Título: Expressão transiente do gene uidA em explantes foliares de Eucalyptus saligna Sm. transformado via Agrobacterium tumefaciens / Transient expression of uidA gene in leaf explants of Eucalyptus saligna Sm. transformed via Agrobacterium tumefaciens
Fonte: Biosci. j. (Online);29(1):1-7, jan./feb. 2013. ilus, tab.
Idioma: pt.
Resumo: O objetivo desse trabalho foi avaliar a influência do pré-cultivo de explantes foliares e do meio de cultura na ressuspensão de Agrobacterium tumefaciens para infecção dos explantes. Os meios MS/2 (50% da concentração de sais) e MS N/2 (50% da concentração de NH4NO3 e KNO3) + PGR (1,0µM de TDZ (thidiazuron) + 0,1 µM de ANA (ácido naftalenoacético)) foram testados na ressuspensão da bactéria para infecção dos explantes. O pré-cultivo consistiu da manutenção dos explantes em meio de cultura para formação de calos (MS N/2 + PGR) durante um dia, sendo o tratamento sem pré-cultivo consistituído dos explantes após a excissão dos mesmos. Os explantes foram mantidos no escuro a 25 ± 2ºC mediante a utilização de plástico preto. O delineamento usado foi o inteiramente casualisado com 20 explantes. Os experimentos foram repetidos duas vezes. O meio MS/2 promoveu resultados superiores (22,4%) comparado ao meio MS N/2 + PGR (14,5%) para a percentagem de área com expressão do gene uidA. Aos 7 dias de cultivo em meio seletivo, a percentagem de área expressando o gene uidA foi 1,6 no MS/2 e 0% para o MS N/2 + PGR. O pré-cultivo produziu resultados superiores aos encontrados sem pré-cultivo, atingindo 31,4% de expressão transiente e no tratamento sem pré-cultivo 2,1%. Após 7 dias de cultivo em meio seletivo, a percentagem de área de expressão dos explantes do tratamento com pré-cultivo permaneceu 4,8% e 0% para o tratamento sem pré-cultivo. Os resultados indicam que o précultivo e ressuspensão da bactéria em meio MS/2 aumentaram a eficiência da expressão transiente do gene uidA em explantes foliares de E. saligna.

The aim of this research was to evaluate the effect of the pre-culture of leaf explants and the effect of the culture medium for the Agrobacterium tumefaciens resuspension to the explant infection. The media, MS/2 (half strength) and MS N/2 (10.3 mM NH4NO3 and 9.4 mM KNO3) + PGR (1.0 µM TDZ (thidiazuron) and 0.1 µM NAA (1-Naphthaleneacetic acid)) were tested for the bacteria resuspension. The pre-culture consisted of the maintenance of the explants on culture medium for callus formation (MS N/2+PGR) during one day and the treatment without pre-culture consisted of the use of the explants after the excision of the same ones. At the end of the co-culture, the MS/2 promoted results superiors to the MS N/2+PGR, and the area percentage that presented expression of the gene uidA was of 22.4% compared at 14.5%. To the 7 days of culture on a medium with kanamycin, the area percentage expressing the gene uidA was 1.6 in MS/2 and 0% for the MS N/2+PGR. At the end of the co-culture, the pre-culture produced results superiors to the found in the treatment without pre-culture, reaching 31.4% of expression and in the treatment without pre-culture 2.1%. After 7 days of culture on a medium with kanamycin, the area percentage of explant expression of the treatment with pre-culture stayed 4.8% and 0% for the treatment without pre-culture. The results indicate that the pre-culture and the bacteria resuspension in MS/2 increase the efficiency of the transient expression of the gene uidA in leaf explants of E. saligna.
Descritores: Transformação Genética
Agrobacterium tumefaciens
Eucalyptus
Genes
Glucuronidase
Responsável: BR396.4



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