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Id: biblio-1045993
Autor: Zeeshan, Nadia; Naz, Saher; Naz, Shumaila; Afroz, Amber; Zahur, Muzna; Zia, Safia.
Título: Heterologous expression and enhanced production of ß-1, 4-glucanase of Bacillus halodurans C-125 in Escherichia coli
Fonte: Electron. j. biotechnol;34:29-36, july. 2018. ilus, tab, graf.
Idioma: en.
Projeto: Higher Education Commission, Pakistan (HEC).
Resumo: Background: Recombinant DNA technology enables us to produce proteins with desired properties and insubstantial amount for industrial applications. Endo-1, 4-ß-glucanases (Egl) is one of the major enzyme involved in degradation of cellulose, an important component of plant cell wall. The present study was aimed at enhancing the production of endo-1, 4-ß-glucanases (Egl) of Bacillus halodurans in Escherichia coli. Results: A putative Egl gene of Bacillus Halodurans was expressed in E. coli by cloning in pET 22b (+). On induction with isopropyl-b-D-1-thiogalactopyranoside, the enzyme expression reached upto ~20% of the cell protein producing 29.2 mg/liter culture. An increase in cell density to 12 in auto-inducing LB medium (absorbance at 600 nm) enhanced ß-glucanase production up to 5.4 fold. The molecular mass of the enzyme was determined to be 39 KDa, which is nearly the same as the calculated value. Protein sequence was analyzed by CDD, Pfam, I TASSER, COACH, PROCHECK Servers and putative amino acids involved in the formation of catalytic, substrate and metal binding domains were identified. Phylogenetic analysis of the ß-glucanases of B. halodurans was performed and position of Egl among other members of the genus Bacillus producing endo-glucanases was determined. Temperature and pH optima of the enzyme were found to be 60°C and 8.0, respectively, under the assay conditions. Conclusion: Production of endo-1, 4 ß-glucanase enzymes from B. halodurans increased several folds when cloned in pET vector and expressed in E. coli. To our knowledge, this is the first report of high-level expression and characterization of an endo-1, 4 ß-glucanases from B. halodurans.
Descritores: Bacillus/enzimologia
Celulases/biossíntese
-Temperatura
Estabilidade Enzimática
Expressão Gênica
Parede Celular/enzimologia
Reação em Cadeia da Polimerase
Clonagem Molecular
Celulases/isolamento & purificação
Celulases/metabolismo
Escherichia coli/metabolismo
Células Vegetais/enzimologia
Concentração de Íons de Hidrogênio
Hidrólise
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1047373
Autor: Shi, Xiaodong; Wu, Yan; Dai, Tingwei; Gu, Yuxi; Wang, Linghui; Qin, Xiaobo; Xu, Ying; Chen, Fang.
Título: JcZFP8, a C2H2 zinc finger protein gene from Jatropha curcas, influences plant development in transgenic tobacco
Fonte: Electron. j. biotechnol;34:76-82, july. 2018. ilus, graf.
Idioma: en.
Projeto: National Key Technology R&D Program of 12th Five-Year Plan of China; . Open Foundation of Key Laboratory of Molecular Genetics, China National Tobacco Corporation (Guizhou Academy of Tobacco Science); . Special Project for Breeding and Cultivation of GMO Varieties of Ministry of Agriculture.
Resumo: Background: Jatropha curcas L., as an important strategic biofuel resource with considerable economic potential, has attracted worldwide attention. However, J. curcas has yet to be domesticated. Plant height, an important agronomic trait of J. curcas, has not been sufficiently improved, and the genetic regulation of this trait in J. curcas is not fully understood. Zinc finger proteins (ZFPs), a class of transcription factors, have previously been shown to play critical roles in regulating multiple aspects of plant growth and development and may accordingly be implicated in the genetic regulation of plant height in J. curcas. Results: In this study, we cloned JcZFP8, a C2H2 ZFP gene in J. curcas. We found that the JcZFP8 protein was localized in the nucleus and contained a conserved QALGGH motif in its C2H2 structure. Furthermore, ectopic expression of JcZFP8 under the control of the 35S promoter in transgenic tobacco resulted in dwarf plants with malformed leaves. However, when JcZFP8 was knocked out, the transgenic tobacco did not show the dwarf phenotype. After treatment with the gibberellic acid (GA) biosynthesis inhibitor paclobutrazol (PAC), the dwarf phenotype was more severe than plants that did not receive the PAC treatment, whereas application of exogenous gibberellin3 (GA3) reduced the dwarf phenotype in transgenic plants. Conclusions: The results of this study indicate that JcZFP8 may play a role in J. curcas plant phenotype through GA-related pathways. Our findings may help us to understand the genetic regulation of plant development in J. curcas and to accelerate breeding progress through engineering of the GA metabolic pathway in this plant. How to cite: Shi X,Wu Y, Dai T, et al. JcZFP8, a C2H2 zinc-finger protein gene from Jatropha curcas, influences plant development in transgenic tobacco.
Descritores: Tabaco/genética
Jatropha
Desenvolvimento Vegetal
Dedos de Zinco CYS2-HIS2/genética
-Reguladores de Crescimento de Plantas/genética
Fatores de Transcrição
Triazóis
Plantas Geneticamente Modificadas/crescimento & desenvolvimento
Clonagem Molecular
Regulação da Expressão Gênica de Plantas
Reação em Cadeia da Polimerase em Tempo Real
Giberelinas
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1047375
Autor: Li, Shanshan; Qin, Kun; Li, Huaying; Guo, Jin; Li, Dejin; Liu, Fang; Tan, Zhilei; Yan, Wei; Qu, Shuling; Zhao, Huabing.
Título: Cloning and characterisation of four catA genes located on the chromosome and large plasmid of Pseudomonas putida ND6
Fonte: Electron. j. biotechnol;34:83-90, july. 2018. tab, ilus, graf.
Idioma: en.
Projeto: Tianjin Research Program of Application Foundation and Advanced Technology; . National Natural Science Foundation of China.
Resumo: Background: Although the functional redundancy of catechol 1,2-dioxygenase (C12O) genes has been reported in several microorganisms, limited enzymes were characterised, let alone the advantage of the coexistence of the multiple copies of C12O genes. Results: In this study, four novel C12O genes, designated catA, catAI, catAII and catAIII, in the naphthalene-degrading strain Pseudomonas putida ND6, were cloned and characterised. Phylogenetic analysis of their deduced amino acid sequences revealed that the four C12O isozymes each formed independent subtrees, together with homologues from other organisms. All four enzymes exhibited maximum activity at pH 7.4 and higher activity in alkaline than in acidic conditions. Furthermore, CatA, CatAI and CatAIII were maximally active at a temperature of 45°C, whereas a higher optimum temperature was observed for CatAII at a temperature of 50°C. CatAI exhibited superior temperature stability compared with the other three C12O isozymes, and kinetic analysis indicated similar enzyme activities for CatA, CatAI and CatAII, whereas that of CatAIII was lower. Significantly, among metal ions tested, only Cu2+ substantially inhibited the activity of these C12O isozymes, thus indicating that they have potential to facilitate bioremediation in environments polluted with aromatics in the presence of metals. Moreover, gene expression analysis at the mRNA level and determination of enzyme activity clearly indicated that the redundancy of the catA genes has increased the levels of C12O. Conclusion: The results clearly imply that the redundancy of catA genes increases the available amount of C12O in P. putida ND6, which would be beneficial for survival in challenging environments.
Descritores: Pseudomonas putida/enzimologia
Pseudomonas putida/genética
Catecol 1,2-Dioxigenase/genética
-Temperatura
Biodegradação Ambiental
Clonagem Molecular
Catecol 1,2-Dioxigenase/análise
Catecol 1,2-Dioxigenase/metabolismo
Genes Bacterianos
Concentração de Íons de Hidrogênio
Isoenzimas
Metais
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1047727
Autor: Nadeem, Muhammad Shahid; Al-Ghamdi, Maryam A; Khan, Jalaluddin Azam; Sadath, Saida; Al-Malki, Abdulaziz.
Título: Recombinant production and biochemical and in silico characterization of lactate dehydrogenase from Geobacillus thermodenitrificans DSM-465
Fonte: Electron. j. biotechnol;35:18-24, sept. 2018. ilus, tab, graf.
Idioma: en.
Resumo: Background: Lactate dehydrogenase (LDH) is an enzyme of glycolytic pathway, ubiquitously found in living organisms. Increased glycolysis and LDH activity are associated with many pathologic conditions including inflammation and cancer, thereby making the enzyme a suitable drug target. Studies on conserved structural and functional domains of LDH from various species reveal novel inhibitory molecules. Our study describes Escherichia coli production and characterization of a moderately thermostable LDH (LDH-GT) from Geobacillus thermodenitrificans DSM-465. An in silico 3D model of recombinant enzyme and molecular docking with a set of potential inhibitors are also described. Results: The recombinant enzyme was overexpressed in E. coli and purified to electrophoretic homogeneity. The molecular weight of the enzyme determined by MALDI-TOF was 34,798.96 Da. It exhibited maximum activity at 65°C and pH 7.5 with a KM value for pyruvate as 45 µM. LDH-GT and human LDH-A have only 35.6% identity in the amino acid sequence. On the contrary, comparison by in silico structural alignment reveals that LDH-GT monomer has approximately 80% identity to that of truncated LDH-A. The amino acids "GEHGD" as well as His179 and His193 in the active site are conserved. Docking studies have shown the binding free energy changes of potential inhibitors with LDH-A and LDH-GT ranging from −407.11 to −127.31 kJ mol−1 . Conclusions: By highlighting the conserved structural and functional domains of LDH from two entirely different species, this study has graded potential inhibitory molecules on the basis of their binding affinities so that they can be applied for in vivo anticancer studies
Descritores: Geobacillus/enzimologia
L-Lactato Desidrogenase/metabolismo
-Simulação por Computador
Estabilidade Enzimática
Reação em Cadeia da Polimerase
Clonagem Molecular
Escherichia coli/metabolismo
Simulação de Acoplamento Molecular
Glicólise
L-Lactato Desidrogenase/genética
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1048187
Autor: Jiménez-Guillen, Doribet; Pérez-Pascual, Daniel; Souza-Perera, Ramón; Godoy-Hernández, Gregorio; Zúñiga-Aguilar, José Juan.
Título: Cloning of the Coffea canephora SERK1 promoter and its molecular analysis during the cell-to-embryo transition
Fonte: Electron. j. biotechnol;36:34-46, nov. 2018. tab, ilus.
Idioma: en.
Projeto: Consejo Nacional de Ciencia y Tecnología; . Comisión Intersecretarial de Bioseguridad de los Organismos Genéticamente Modificados; . DJG and DPP received CONACYT Ph.D..
Resumo: Background: Somatic embryogenesis receptor-like kinase 1 (SERK1) is a cell membrane receptor active in different plant tissues and involved in cell differentiation activities including somatic embryogenesis. The identification of promoter elements responsible for SERK1 expression during the onset of somatic embryogenesis can be useful to understand the molecular regulation of the cell-to embryo transition, and these promoter elements represent biotechnological tools in plant organ tissue culture. Results: A −1,620 bp DNA sequence located upstream of the Coffea canephora SERK1 gene homologue (CcSERK1) was isolated, and then, different segments containing key response elements (REs) for somatic embryogenesis onset and development were fused to the uidA (encoding a ß-glucuronidase, GUS) reporter gene to evaluate its expression in transgenic leaf explants. DNA segments of −1,620 and −1048 bp in length directed uidA expression with patterns in leaf explants similar to those occurring during somatic embryogenesis. When a −792-bp fragment was used, uidA expression disappeared only in leaf explants and pro-embryogenic mass but persisted in developing embryos. No uidA expression was detected in any embryogenic stage when a −618-bp fragment was used. Conclusion: DNA deletions showed that a −1048-bp sequence located upstream of the CcSERK1 gene is sufficient to direct gene expression during the onset and the development of C. canephora somatic embryogenesis. The DNA segment located between −1048 and −792 bp (containing BBM and WUS REs) is needed for gene expression before embryogenesis onset but not during embryo development. The promoter segment between −792 and −618 bp (including GATA, ARR1AT, and ANT REs) regulates gene expression in developing embryos.
Descritores: Proteínas de Plantas/genética
Proteínas Quinases/genética
Coffea/genética
-Biotecnologia
Expressão Gênica
Regiões Promotoras Genéticas
Plantas Geneticamente Modificadas
Clonagem Molecular
Genes Reporter
Regulação da Expressão Gênica de Plantas
Desenvolvimento Embrionário
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1047976
Autor: Yang, Xiulian; Ding, Wenjie; Yue, Yuanzheng; Xu, Chen; Wang, Xi; Wang, Lianggui.
Título: Cloning and expression analysis of three critical triterpenoid pathway genes in Osmanthus fragrans
Fonte: Electron. j. biotechnol;36:1-8, nov. 2018. ilus, graf.
Idioma: en.
Projeto: Application Demonstration of the Key Technology and Innovation in Breeding the Six Precious Colored Tree Species of Jiangsu Province; . Academic Programs Project of Jiangsu Higher Education Institutions.
Resumo: Background: Osmanthus fragrans is an important ornamental tree and has been widely planted in China because of its pleasant aroma, which is mainly due to terpenes. The monoterpenoid and sesquiterpenoid metabolic pathways of sweet osmanthus have been well studied. However, these studies were mainly focused on volatile small molecule compounds. The molecular regulation mechanism of synthesis of large molecule compounds (triterpenoids) remains unclear. Squalene synthase (SQS), squalene epoxidase (SQE), and beta-amyrin synthase (BETA-AS) are three critical enzymes of the triterpenoid biosynthesis pathway. Results: In this study, the full-length cDNA and gDNA sequences of OfSQS, OfSQE, and OfBETA-AS were isolated from sweet osmanthus. Phylogenetic analysis suggested that OfSQS and OfSQE had the closest relationship with Sesamum indicum, and OfBETA-AS sequence shared the highest similarity of 99% with that of Olea europaea. The qRT-PCR analysis revealed that the three genes were highly expressed in flowers, especially OfSQE and OfBETA-AS, which were predominantly expressed in the flowers of both "Boye" and "Rixiang" cultivars, suggesting that they might play important roles in the accumulation of triterpenoids in flowers of O. fragrans. Furthermore, the expression of OfBETA-AS in the two cultivars was significantly different during all the five flowering stages; this suggested that OfBETA-AS may be the critical gene for the differences in the accumulation of triterpenoids. Conclusion: The evidence indicates that OfBETA-AS could be the key gene in the triterpenoid synthesis pathway, and it could also be used as a critical gene resource in the synthesis of essential oils by using bioengineered bacteria.
Descritores: Triterpenos/metabolismo
Clonagem Molecular
Oleaceae/genética
-Farnesil-Difosfato Farnesiltransferase/metabolismo
Óleos Voláteis
Expressão Gênica
Reação em Cadeia da Polimerase
Oleaceae/enzimologia
Esqualeno Mono-Oxigenase/metabolismo
Odorantes
Responsável: CL1.1 - Biblioteca Central


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Texto completo SciELO Chile
Texto completo
Id: biblio-950778
Autor: Muzaffar, Adnan; Kiani, Sarfraz; Khan, Muhammad Azmat Ullah; Rao, Abdul Qayyum; Ali, Arfan; Awan, Mudassar Fareed; Iqbal, Adnan; Nasir, Idrees Ahmad; Shahid, Ahmad Ali; Husnain, Tayyab.
Título: Chloroplast localization of Cry1Ac and Cry2A protein- an alternative way of insect control in cotton
Fonte: Biol. Res;48:1-11, 2015. ilus, tab.
Idioma: en.
Resumo: BACKGROUND: Insects have developed resistance against Bt-transgenic plants. A multi-barrier defense system to weaken their resistance development is now necessary. One such approach is to use fusion protein genes to increase resistance in plants by introducing more Bt genes in combination. The locating the target protein at the point of insect attack will be more effective. It will not mean that the non-green parts of the plants are free of toxic proteins, but it will inflict more damage on the insects because they are at maximum activity in the green parts of plants. RESULTS: Successful cloning was achieved by the amplification of Cry2A, Cry1Ac, and a transit peptide. The appropriate polymerase chain reaction amplification and digested products confirmed that Cry1Ac and Cry2A were successfully cloned in the correct orientation. The appearance of a blue color in sections of infiltrated leaves after 72 hours confirmed the successful expression of the construct in the plant expression system. The overall transformation efficiency was calculated to be 0.7%. The amplification of Cry1Ac-Cry2A and Tp2 showed the successful integration of target genes into the genome of cotton plants. A maximum of 0.673 µg/g tissue of Cry1Ac and 0.568 µg/g tissue of Cry2A was observed in transgenic plants. We obtained 100% mortality in the target insect after 72 hours of feeding the 2nd instar larvae with transgenic plants. The appearance of a yellow color in transgenic cross sections, while absent in the control, through phase contrast microscopy indicated chloroplast localization of the target protein. CONCLUSION: Locating the target protein at the point of insect attack increases insect mortality when compared with that of other transgenic plants. The results of this study will also be of great value from a biosafety point of view.
Descritores: Proteínas de Bactérias/genética
Proteínas Recombinantes de Fusão
Cloroplastos/genética
Controle de Insetos/métodos
Gossypium/genética
Endotoxinas/genética
Proteínas Hemolisinas/genética
Lepidópteros
-Bacillus thuringiensis
Proteínas de Bactérias/análise
Resistência a Inseticidas/genética
Imuno-Histoquímica
Expressão Gênica/genética
Cloroplastos/metabolismo
Reação em Cadeia da Polimerase
Microscopia de Contraste de Fase
Plantas Geneticamente Modificadas
Clonagem Molecular
Primers do DNA
Folhas de Planta/genética
Transgenes/fisiologia
Endotoxinas/análise
Fusão Gênica
Proteínas Hemolisinas/análise
Inseticidas
Larva
Limites: Animais
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: CL1.1 - Biblioteca Central


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Texto completo SciELO Brasil
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Id: biblio-1132217
Autor: Jamal-Livani, Narges; Nikokar, Elham; Safdari, Yaghoub.
Título: In-Situ Gel-Free Plasmid Reassembling for Rapid Gene Subcloning and Truncation
Fonte: Braz. arch. biol. technol;63:e20190223, 2020. tab, graf.
Idioma: en.
Projeto: Golestan University of Medical Sciences.
Resumo: Abstract Gene subcloning, a process in which the nucleotide sequence of interest is excised from on plasmid and inserted into another, seems to be an easy task to done. However, not all subcloning attempts are successful, even when the insert sequence and the double digested target plasmid are successfully purified form agarose gel and thought to be ready for subsequent ligation. In the current study we introduce a reliable, easy, and time consuming method for gene subcloning and also truncation. The stages are all carried out in a single microtube without any running on a gel, making it possible to accomplish a successful gene subcloning or truncation even with low concentrations of DNA molecules. Summarily, subcloning is achieved by mixing the plasmids of interest in a microtube and subjecting to restriction enzymes whose restriction sites flank the segment that is going to be subcloned. Digestion mixture is precipitated in the same microtube using isopropanol and the resultant DNA molecules are allowed to take part in a ligation reaction. The recombinant plasmids of interest are screened by colony PCR. Truncation is achieved by double- digestion of the plasmid of interest using a restriction enzyme whose restriction site flanks the segment that is going to be cut out.
Descritores: Plasmídeos/genética
Clonagem Molecular/métodos
Vetores Genéticos
-Reação em Cadeia da Polimerase
Responsável: BR1.1 - BIREME


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Id: lil-756701
Autor: Ferreira, José Roberto de Oliveira.
Título: Caracterização funcional de variantes de patogenicidade incerta, MLH1 LEU676PRO E MSH2 MET729ILE, encontradas em pacientes diagnosticados com síndrome de Lynch / Functional characterization of variants of uncertain pathogenicity, MLH1 Leu676Pro and MSH2 Met729Ile, found in patients diagnosed with Lynch syndromeneoplasias and uterine cervix squamous cell carcinoma.
Fonte: São Paulo; s.n; 2014. 110 p. tab, ilus, quadros.
Idioma: pt.
Tese: Apresentada a Fundação Antônio Prudente para obtenção do grau de Doutor.
Resumo: O melanoma cutâneo é uma neoplasia que acomete indivíduos jovens e apresenta comportamento agressivo quando diagnosticado tardiamente. Sendo assim, novos métodos diagnósticos auxiliares ao exame clínico, como a dermatoscopia e a microscopia confocal in vivo (MC), têm sido desenvolvidos com o objetivo de melhorar a acurácia diagnóstica desse tumor. Semelhante à dermatoscopia, a MC revela detalhes morfológicos da arquitetura tecidual no plano paralelo à pele e, além disso, fornece imagens instantâneas com alta magnificação e resolução celular. A realização de cortes histológicos transversais (mesmo plano da dermatoscopia e MC) poderia contribuir para melhor caracterizar os achados observados tanto na dermatoscopia quanto na MC. Não existem relatos na literatura médica comparando as características dermatoscópicas, a MC e os achados histopatológicos em cortes transversais. O objetivo deste estudo foi descrever a técnica para realização dos cortes histológicos transversais e comparar as principais características dermatoscópicas do melanoma cutâneo à MC e à histopatologia em cortes perpendiculares e transversais, no intuito de oferecer uma interpretação mais precisa dos achados celulares e arquiteturais observados in vivo. Foram avaliadas 65 lesões com diagnóstico dermatoscópico de melanoma cutâneo de 63 pacientes recrutados no Núcleo de Câncer de Pele e Dermatologia do A.C. Camargo Cancer Center no período de junho de 2011 a abril de 2013. Uma forma fácil, segura e confiável para a realização dos cortes histológicos transversais foi apresentada. Os aspectos celulares e arquiteturais no exame de MC das principais características dermatoscópicas do melanoma cutâneo foram determinados e comparados aos achados histopatológicos nos cortes transversais e perpendiculares. A MC permitiu a identificação de uma nova estrutura chamada de “papila em mitocôndria” que pode representar um critério adicional para o diagnóstico do melanoma in situ. Os cortes histológicos...

Cutaneous melanoma is a cancer that affects young individuals and shows aggressive behavior when diagnosed lately. Therefore, new diagnostic tools to unaided eye, such as dermoscopy and in vivo reflectance confocal microscopy (RCM), have been developed with the aim of improving the diagnostic accuracy of this tumor. Similarly to dermoscopy, RCM reveals morphological details of tissue architecture in parallel plane to the skin and, moreover, provides instant images with high magnification and cellular level resolution. The performance of transverse histopathological sections (same plane of dermoscopy and RCM) could help to better characterize the features observed in both dermoscopy and RCM. There are no reports in the medical literature comparing dermoscopic, RCM and histopathological features in transverse sections. The purpose of this study was to describe the technique for acquiring the transverse sections and compare the main dermoscopic features of cutaneous melanoma to the RCM and histopathology in perpendicular and transverse sections, in order to offer a more precise interpretation of the cellular and architectural features observed in vivo. This study included 65 lesions with dermoscopic diagnosis of cutaneous melanoma in 63 patients recruited at the Dermatology Center of the AC Camargo Cancer Center from June 2011 to April 2013. An easy, safe and reliable way for handle the transverse sections was presented. The RCM cellular and architectural aspects of the main melanoma dermoscopic features were determined and compared to histopathological findings in the transverse and perpendicular sections. We described a new structure called “papillae in mitochondria” which may represent an additional clue for the melanoma in situ diagnosis. The transverse sections allowed a more precise interpretation of the main RCM features in cutaneous melanoma...
Descritores: Clonagem Molecular
Imunofluorescência
Neoplasias Colorretais Hereditárias sem Polipose
Reparo do DNA
Limites: Humanos
Responsável: BR30.1 - Biblioteca
BR30.1


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Texto completo SciELO Chile
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Id: biblio-1011422
Autor: Wei, Fan; Tang, Danfeng; Li, Zengqiang; Kashif, Muhammad Haneef; Khan, Aziz; Lu, Hai; Jia, Ruixing; Chen, Peng.
Título: Molecular cloning and subcellular localization of six HDACs and their roles in response to salt and drought stress in kenaf (Hibiscus cannabinus L)
Fonte: Biol. Res;52:20, 2019. tab, graf.
Idioma: en.
Projeto: National Natural Science Foundation of China; . Modern Agro-industry Technology Research System.
Resumo: BACKGROUND: Histone acetylation is an important epigenetic modification that regulates gene activity in response to stress. Histone acetylation levels are reversibly regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). The imperative roles of HDACs in gene transcription, transcriptional regulation, growth and responses to stressful environment have been widely investigated in Arabidopsis. However, data regarding HDACs in kenaf crop has not been disclosed yet. RESULTS: In this study, six HDACs genes (HcHDA2, HcHDA6, HcHDA8, HcHDA9, HcHDA19, and HcSRT2) were isolated and characterized. Phylogenetic tree revealed that these HcHDACs shared high degree of sequence homology with those of Gossypium arboreum. Subcellular localization analysis showed that GFP-tagged HcHDA2 and HcHDA8 were predominantly localized in the nucleus, HcHDA6 and HcHDA19 in nucleus and cytosol. The HcHDA9 was found in both nucleus and plasma membranes. Real-time quantitative PCR showed that the six HcHDACs genes were expressed with distinct expression patterns across plant tissues. Furthermore, we determined differential accumulation of HcHDACs transcripts under salt and drought treatments, indicating that these enzymes may participate in the biological process under stress in kenaf. Finally, we showed that the levels of histone H3 and H4 acetylation were modulated by salt and drought stress in kenaf. CONCLUSIONS: We have isolated and characterized six HDACs genes from kenaf. These data showed that HDACs are imperative players for growth and development as well abiotic stress responses in kenaf.
Descritores: Estresse Fisiológico/fisiologia
Hibiscus/enzimologia
Histona Acetiltransferases/fisiologia
Secas
Histona Desacetilases/fisiologia
-Ativação Transcricional/fisiologia
Clonagem Molecular
Hibiscus/crescimento & desenvolvimento
Hibiscus/fisiologia
Reação em Cadeia da Polimerase em Tempo Real
Responsável: CL1.1 - Biblioteca Central



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