Estudo do Cultivo de dois clones de Escherichia coli recombinantes (eIF, LACK) para a Expressão de Antígenos da Leishmania chagasi

With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and identification of new proteins, mainly with therapeutical purposes. The Escherichia coli has been procarioto predominant in the studies of genetic engineering due to wealth of information regarding its metabolism. Despite the expressivo advance of the studies of molecular biology and the immunology of the infections, it does not exist, currently, no prophylactic drug capable to prevent calazar. Of this form, it exists a great necessity of specific antigen identification for the vaccine development and kits for disgnostic against the visceral Leishmaniose. In this context, this work objectified to study the recombinant antigen expression of the Leishmania chagasi during the culture of Escherichia coli in shaker. A first set of assays was carried through with the objective of if knowing the kinetic behavior of the growth of two clones recombinant proteins (eIF, LACK) in two different compositions of culture medium (2xTY, TB) supplemented by antibiotics, without IPTG addition. In the second stage of the assays, the procedure of induction for IPTG was carried through, in order to verify the influence of the composition of the ways tested in the expression them recombinant proteins. On the basis of the gotten results, can be observed that the high complexity of culture medium favored the kinetic one of growth of clones recombinant (eIF, LACK), however, to if to deal with the assays submitted to the procedure of induction for IPTG, the raised complexity of culture medium did not favor the expression of recombinant proteins. On the other hand, they had been gotten resulted positive for all clones recombinant (eIF, LACK) tested, confirmed through the eletroforético profile

Alteração fisiológica da cana-de-açúcar pela aplicação de Glyphosate e Sulfumeturon-Methyl

O bloqueio parcial das rotas onde atuam os herbicidas, com uso de baixas doses, pode ter implicações importantes, como a alteração do balanço de processos metabólicos nas plantas. Assim, foi conduzido no ano agrícola 2006/2007 um experimento em cana-soca de segundo corte na Fazenda Jurema, pertencente ao grupo COSAN, município de Barra Bonita-SP, para verificar os efeitos do glyphosate e do sulfumeturon-methyl, em subdoses, no comportamento fisiológico da cana-de-açúcar pelos níveis de clorofila e carotenoides. Os tratamentos constituíram-se da aplicação de dois herbicidas: sulfumeturon-methyl (Curavial 360 e.a. kg-1) e glyphosate (Roundup 480 i.a. kg-1), isolados e em misturas, em diferentes doses, e um tratamento controle, sem a aplicação dos herbicidas. As doses utilizadas foram: glyphosate 200 mL p.c. ha-1; glyphosate 400 mL p.c. ha -1; glyphosate 200 mL p.c. ha -1 + 10 g p.c. ha-1 de sulfumeturon-methyl; glyphosate 150 mL p.c. ha -1 + 12 g p.c. ha -1 de sulfumeturon-methyl; e sulfumeturon-methyl 20 g p.c. ha -1. O delineamento experimental utilizado foi o de blocos casualizados, com quatro repetições. As avaliações foram realizadas 15 e 30 dias após o plantio (DAP) e 30, 60, 90, 120, 150 e 180 dias após a colheita (DAC). As folhas foram cortadas padronizando-se o mesmo peso e área foliar. Para determinação do conteúdo de clorofila e carotenoides, amostras de 0,2 g de tecido foliar fresco foram preparadas e os extratos filtrados, sendo efetuadas leituras em espectrofotômetro (663 e 645 nm para clorofilas a e b, respectivamente). A aplicação de glyphosate e sulfumeturon-methyl nas maiores doses interferiu no conteúdo de carotenoides quando estes foram comparados com a testemunha. A maior dose de glyphosate diminuiu significativamente o conteúdo de clorofilas e carotenoides na cana-de-açúcar, porém esse resultado não se manteve quando a dose foi reduzida para 200 mL p.c. ha-1 . Os teores de clorofila foram inversamente proporcionais aos níveis Fe. A aplicação de sulfumeturon-methyl não interferiu nos teores de clorofila, no entanto os níveis de carotenoides se mostraram mais sensíveis e seus teores reduzidos. As alterações observadas nos níveis de clorofilas e carotenoides pela aplicação dos produtos podem afetar de maneira distinta o metabolismo da fotossíntese pela absorção e/ou conversão de energia.

Teores de lignina e celulose em plantas de cana-de-açúcar em função da aplicação de maturadores

O objetivo deste trabalho foi avaliar os teores de lignina e celulose em plantas de cana-de-açúcar após a aplicação de dois maturadores para a colheita. O experimento foi conduzido em uma área de cana-soca, cultivar SP 803280, no município de Igaraçu do Tietê/SP. O delineamento experimental utilizado foi o de blocos casualizados, com quatro repetições. Os tratamentos constituíram-se da aplicação de dois maturadores: sulfometuron-methyl (Curavial) e glyphosate (Roundup original). As doses utilizadas foram: glyphosate a 72 g e.a. ha-1; glyphosate a 144 g e.a. ha-1 ; glyphosate a 72 g e.a. ha-1 + sulfometuron methyl a 10 g p.c. ha-1; glyphosate a 108 g e.a. ha-1 + sulfometuron-methyl a 12 g p.c. ha-1; sulfometuron-methyl a 20 g p.c. ha-1; e a testemunha sem aplicação de maturadores. As análises de lignina e celulose foram realizadas pelo método lignina em detergente ácido modificado. O glyphosate e o sulfometuron-methyl alteraram os níveis de lignina no momento da colheita, e esse efeito foi observado também durante o crescimento da cana-de-açúcar (meses após a aplicação desses produtos). O glyphosate a 72 g e.a. ha-1 promoveu reduções nos teores de lignina, na colheita e durante o crescimento da cana-de-açúcar, quando comparados com os da testemunha, enquanto o sulfometuron-methyl isolado na menor dose (10 g ha-1) promoveu aumento nos teores desse biopolímero na soqueira da cana-de-açúcar.

Avaliação, identificação e atividade enzimática de bactérias psicotróficas presentes no leite cru refrigerado

Extended storage of refrigerated milk can lead to reduced quality of raw and processed milk, which is a consequence of the growth and metabolic activities of psychrotrophic bacteria, able to grow under 7oC or lower temperatures. Although most of these microorganisms are destroyed by heat treatment, some have the potential to produce termoresistant proteolytic and lipolytic enzymes that can survive even UHT processing and reduce the processed products quality. Recently, the IN 51 determineds that milk should be refrigerated and stored at the farm what increased the importance of this group of microorganisms. In this work, psychrotrophic bacteria were isolated from 20 communitarian bulk tanks and 23 individual bulk tanks from dairy farms located at Zona da Mata region of Minas Gerais State and from southeastern Rio de Janeiro. Selected milk dilutions were plated on standard agar and after incubation for 10 days at 7oC, five colonies were isolated, firstly using nutrient agar and after using McConkey agar for 24 hours at 21oC. The isolates were identified by morphology, Gram stain method, catalase production, fermentative/oxidative metabolism and by API 20E, API 20NE, API Staph, API Coryne or API 50 CH (BioMerieux). In order to ensure reproductibility, API was repeated for 50% of the isolates. Species identification was considered when APILAB indexes reached 75% or higher. 309 strains were isolated, 250 Gram negative and 59 Gram positive. 250 Gram negative isolates were identified as: Acinetobacter spp. (39), Aeromonas spp. (07), A. Hydrophila (16), A. sobria (1), A. caviae (1), Alcaligenes feacalis (1), Burkholderia cepacia (12), Chryseomonas luteola (3), Enterobacter sp. (1), Ewingella americana(6), Hafnia alvei (7), Klebsiella sp. (1), Klebsiella oxytoca (10), Yersinia spp. (2), Methylobacterium mesophilicum (1), Moraxella spp. (4), Pantoea spp. (16), Pasteurella sp. (1), Pseudomonas spp. (10), P. fluorescens (94), P. putida (3), Serratia spp. (3), Sphigomonas paucomobilis (1). Five isolates kept unidentified. Pseudomonas was the predominant bacteria found (43%) and P. fluorescens the predominant species (37.6%), in accordance with previous reports. Qualitative analysis of proteolytic and lipolytic activity was based on halo formation using caseinate agar and tributirina agar during 72 hours at 21oC and during 10 days at 4°C, 10oC and 7°C. Among 250 Gram negative bacteria found, 104 were identified as Pseudomonas spp. and 60,57% of this group showed proteolytic and lipolytic acitivities over all four studied temperatures. 20% of Acinetobacter, Aeromonas, Alcaligenes, Burkholderia, Chryseomonas, Methylobacterium, Moraxella presented only lipolytic activity. Some isolates presented enzymatic activity in one or more studied temperatures. Among Gram positive bacteria, 30.51% were proteolytic and lipolytic at 10oC, 8.47% were proteolytic at 7oC, 10oC, and 21oC, 8.47% were proteolytic at all studied temperatures (4oC, 7oC, 10oC and 21oC) and 3.38% were proteolytic only at 21oC. At 4oC, only one isolate showed proteolytic activity and six isolates were lipolytic. In relation to Gram negative microorganisms, 4% were proteolytic and lipolytic at 7oC, 10oC and 21oC, 10% were proteolytic at 10oC and 4.4% were lipolytic at 4oC, 7oC, 10oC and 21oC, while 6.4% of all isolates were proteolytic and lipolytic at 10oC and 21oC as well as lipolytic at 4oC and 7oC. These findings are in accordance with previous researches that pointed out Pseudomonas as the predominant psycrotrophic flora in stored refrigerated raw milk

Qualidade pós-colheita de goiabas 'Branca de Kumagai', tratadas com cloreto de cálcio

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Monitoramento em tempo real de processos fermentativos por espectroscopia no infravermelho próximo (NIRS)

A chemical process optimization and control is strongly correlated with the quantity of information can be obtained from the system. In biotechnological processes, where the transforming agent is a cell, many variables can interfere in the process, leading to changes in the microorganism metabolism and affecting the quantity and quality of final product. Therefore, the continuously monitoring of the variables that interfere in the bioprocess, is crucial to be able to act on certain variables of the system, keeping it under desirable operational conditions and control. In general, during a fermentation process, the analysis of important parameters such as substrate, product and cells concentration, is done off-line, requiring sampling, pretreatment and analytical procedures. Therefore, this steps require a significant run time and the use of high purity chemical reagents to be done. In order to implement a real time monitoring system for a benchtop bioreactor, these study was conducted in two steps: (i) The development of a software that presents a communication interface between bioreactor and computer based on data acquisition and process variables data recording, that are pH, temperature, dissolved oxygen, level, foam level, agitation frequency and the input setpoints of the operational parameters of the bioreactor control unit; (ii) The development of an analytical method using near-infrared spectroscopy (NIRS) in order to enable substrate, products and cells concentration monitoring during a fermentation process for ethanol production using the yeast Saccharomyces cerevisiae. Three fermentation runs were conducted (F1, F2 and F3) that were monitored by NIRS and subsequent sampling for analytical characterization. The data obtained were used for calibration and validation, where pre-treatments combined or not with smoothing filters were applied to spectrum data. The most satisfactory results were obtained when the calibration models were constructed from real samples of culture medium removed from the fermentation assays F1, F2 and F3, showing that the analytical method based on NIRS can be used as a fast and effective method to quantify cells, substrate and products concentration what enables the implementation of insitu real time monitoring of fermentation processes

Análise de fluxos metabólicos para otimização da síntese do antibiótico cosmomicina por Streptomyces olindensis ICB20

Metabolic flux analysis (MFA) is a powerful tool for analyzing cellular metabolism. In order to control the growth conditions of a specific organism, it is important to have a complete understanding of its MFA. This would allowed us to improve the processes for obtaining products of interest to human and also to understand how to manipulate the genome of a cell, allowing optimization process for genetic engineering. Streptomyces olindensis ICB20 is a promising producer of the antibiotic cosmomycin, a powerful antitumor drug. Several Brazilian researchers groups have been developing studies in order to optimize cosmomycin production in bioreactors. However, to the best of our knowledge, nothing has been done on metabolic fluxes analysis field. Therefore, the aim of this work is to identify several factors that can affect the metabolism of Streptomyces olindensis ICB20, through the metabolic flux analysis. As a result, the production of the secondary metabolite, cosmomycin, can be increased. To achieve this goal, a metabolic model was developed which simulates a distribution of internal cellular fluxes based on the knowledge of metabolic pathways, its interconnections, as well as the constraints of microorganism under study. The validity of the proposed model was verified by comparing the computational data obtained by the model with the experimental data obtained from the literature. Based on the analysis of intracellular fluxes, obtained by the model, an optimal culture medium was proposed. In addition, some key points of the metabolism of Streptomyces olindensis were identified, aiming to direct its metabolism to a greater cosmomycin production. In this sense it was found that by increasing the concentration of yeast extract, the culture medium could be optimized. Furthermore, the inhibition of the biosynthesis of fatty acids was found to be a interesting strategy for genetic manipulation. Based on the metabolic model, one of the optimized medium conditions was experimentally tested in order to demonstrate in vitro what was obtained in silico. It was found that by increasing the concentration of yeast extract in the culture medium would induce to an increase of the cosmomycin production

Absorção e translocação de fósforo em dois cultivares de mamoneira (Ricinus communis L.), 'Campinas' e 'Guarani', com uso de 32p

Com o objetivo de estudar a absorção e translocação do fósforo na cultura da mamoneira, instalou-se um ensaio em vasos com os cultivares 'Campinas' e 'Guarani' e empregou-se o fósforo radioativo (32p) como traçador. Os quatro tratamentos constaram da aplicação ao solo de 65 ppm de P contendo 32p, quando as plantas apresentavam seis, nove, doze e quatorze folhas, respectivamente e cortadas com nove, doze e quatorze folhas e as do quarto tratamento após 21 días da aplicação do fertilizante marcado. Através dos dados obtidos, pode-se concluir que, neste solo com baixo teor de fósforo, a aplicação deste elemento, na época da emissão de ínflorescências primárias, não influi no desenvolvimento das inflorescências, uma vez que se formam as expensas de fósforo previamente acumulado no pecíolo e no limbo, e que para lá se desloca a fim de participar do metabolismo de formação e desenvolvimento. em outros termos, o fósforo, quando aplicado tardiamente, não incrementará a produção dos cachos primários e, consequentemente, não pode esperar que o potencial máximo do cultivar seja atingido, pois os cachos primários representam, no mínimo, 30% da produção total da mamoneira.

Os reservatórios eutrofizados da região tropical semiárida atuam como emissores ou como sequestradores de dióxido de carbono?

The aquatic ecosystems can play a role as carbon-dioxide-source or carbon-dioxide-sink systems due to the high predominance of heterotrophic or autotrophic metabolism. The primary production can strongly affect the carbon balance (CO2) through the consumption of carbon dioxide in the photosynthesis, especially in eutrophic environment, acting as a carbon sink. The present study tested the hypothesis that the eutrophic reservoirs in tropical semi-arid region are carbon dioxide-sink systems due to the high primary productivity presented in these systems. Five Brazilian reservoirs from the semi-arid in the northeast region were monitored monthly during four years (2010 to 2013) with a prolonged drought event identified during the study. The results showed an increasing level of eutrophication over the period of prolonged drought, with the predominance of autotrophy. Significant negative correlations were observed between the partial pressure of CO2 (pCO2) (p<0,001) and chlorophyll-a in the Boqueirão, Passagem das Traíras, Dourado and Gargalheiras reservoirs, showing a pattern of the carbon dioxide-sink systems. However, this pattern was not found in Cruzeta reservoir. In summary, in the tropical semi-arid region, hydrological and morphometric variables can lead to different behaviors of the water-supply reservoirs on the carbon metabolism. The eutrophic reservoirs evaluated showed a negative relationship between pCO2 and Chl-a, which suggests that these water bodies show an autotrophic metabolism and behave as carbon dioxide- sink systems

Respostas lactacidêmicas de ratos ao treinamento intermitente de alta intensidade

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comportamento da produção de espécies reativas de oxigênio em miocárdio de ratos submetidos a treinamento de baixa intensidade em diferentes temperaturas

INTRODUÇÃO: A prática de exercício físico proporciona aumento da produção de espécies reativas de oxigênio (ERO) resultantes do metabolismo aeróbio e, gera uma quantidade significativa de calor, em conseqüência da produção de energia, resultando em sobrecarga orgânica. A associação entre ERO e exercício, e entre exercício e variações da temperatura ambiente têm sido estudadas, contudo, há escassez de informações que considere a associação entre produção de radicais livres no miocárdio e atividade física em temperatura elevada. OBJETIVO: Comparar a produção de ERO em miocárdio de ratos submetidos ao treinamento de baixa intensidade em diferentes temperaturas. MÉTODOS: Foram utilizados 20 ratos Wistar, machos, jovens, peso (250 a 280g), divididos em quatro grupos: G1 (n = 5) expostos ao treinamento e calor (39º ± 1C); G2 (n = 5) expostos somente ao calor durante o mesmo período de G1, sem treinamento; G3 (n = 5) expostos ao treinamento em temperatura ambiente (22º ± 1C); G4 (n = 5) expostos à temperatura ambiente sem treinamento. O treinamento foi realizado em esteira rolante climatizada por cinco semanas, evoluindo 5 minutos a cada duas sessões finalizando em 60 minutos em baixa intensidade 8m/min. O ambiente foi controlado entre 39 ± 1ºC e 22 ± 1ºC e entre 40 e 60 % de umidade relativa. A lipoperoxidação foi avaliada por Quimiluminescência (QL). A análise dos dados foi realizada a partir do teste Two Way ANOVA para análise da QL e t de student para a Capacidade Antioxidante Total (TRAP). RESULTADOS: A análise da QL revelou uma curva de emissão de luz significantemente mais baixa para o grupo exposto ao exercício em normotermia comparado aos sedentários mantidos no calor. A análise da TRAP mostrou diminuição em todos os grupos experimentais em relação ao G4. CONCLUSÃO: Concluiu-se que houve níveis menores de produção de ERO nos grupos submetidos somente ao calor ou somente ao exercício.

Adaptabilidade de híbridos entre Gossypium barbadense E G. hirsutum N contendo o gene cry1Ac

Hybrids among transgenic plants and related species are expected to occur if they are sympatric and when there are not crossing barriers; as is the case, in Brazil, of cry1Ac transgenic cotton and Gossypium barbadense. This species has been maintained as dooryard plants, and should be preserved as a genetic resource. Hybrids were evaluated about traits related to fitness, leading to infer about its chances of survivor and selection. A barbadense genotype collected at the state of Mato Grosso was outcrossed to the variety DP 404, containing the gene cry1Ac, and to the isoline DP 404. All the F1 individuals and 122 among 170 F2 individuals expressed the toxin, and presented levels of resistance to pink bollworm (Pectinophora gossypiella) and cotton leafworm (Alabama argillacea) equivalent to the transgenic parent and superior to the isoline, barbadense or non transgenic hybrids. The percentage of germination and number of days to germinate did not differ among genotypes. Anthesis of the first flower and opening of the first cotton boll occurred earlier for herbaceous cotton and F1 hybrids than F2 population in average; all the populations presented a number of days to flower and opening of the first boll smaller then barbadense. The highest plants were barbadenses, and herbaceus the smallest, with F1 and F2 populations presenting intermediary heights. The number of seeds per plants were superior for F1 hybrids an herbaceous cotton, F2 populations were in average intermediary; the barbadense genotype produced the smallest number of seeds per plant. Pink bollworm, mainly, and also cotton leafworm, are important barbadense pests, so the transgene positive effect could favor the selection of hybrids, and hence G. hirsutum genome, against the maintenance of pure G. barbadense genome. The selection may be influenced by the plant uses: the smaller size of hybrids when compared to the barbadense may lead them to be differentiated from these parents to which medicinal properties are attributed; on the other hand, the greater boll production may favor hybrids maintenance with the purpose of producing lamp wicks, or use as an ornamental or swab

Prospecção de genes de interesse biotecnológico : uma abordagem metagenômica

The total number of prokaryotic cells on Earth has been estimated at 4 to 6x1030 and only about 1% of microorganisms present in the environment can be cultivated by standard techniques of cultivation and plating. Therefore, it is a huge biological and genetic pool that can be exploited, for the identification and characterization of genes with biotechnological potential. Within this perspective, the metagenomics approach was applied in this work. Functional screening methods were performed aiming to identify new genes related to DNA repair and / or oxidative stress resistance, hydrocarbon degradation and hydrolytic activities (lipase, amylase and protease). Metagenomic libraries were built utilizing DNA extracted from soil samples collected in João Câmara RN. The libraries were analyzed functionally using specific substrate containing solid medium (hydrolytic activity), supplemented with H2O2 (DNA repair and / or resistance to oxidative stress) and liquid medium supplemented with light Arabian oil (activity, degradation of hydrocarbons). After confirmation of activity and exclusion of false-positive results, 49 clones were obtained, being 2 positive for amylase activity, 22 resistant to oxidative stress generated by H2O2 and 25 clones active for hydrocarbons degradation. Analysis of the sequences showed hypothetical proteins, dienelactona hydrolase, DNA polymerase, acetyltransferase, phosphotransferase, methyltransferase, endonucleases, among other proteins. The sequence data obtained matched with the functions tested, highlighting the success of metagenomics approaches combined with functional screening methods, leading to very promising results

Caracterização in situ e diversidade genética de algodoeiros mocós (Gossypium hissutum raça marie galante) da Região Nordeste do Brasil

Brazil is one of the major centers of diversity for polyploid cotton plants; these plants belong to the genus Gossypium, which has three known species: G. hirsutum, G. barbadense and G. mustelinum. The Northeast is the only region where the three species occur, the last group being endemic. Northeast s cotton plants can be important sources of variability for genetic breeding. It is believed that great part of local diversity is being lost, due to economic, political, cultural and agricultural problems. In an attempt to mitigate this loss and delineate conservation strategies it is necessary to know how the species are found where they occur. The objective was to characterize and determine how plants are maintained in situ in the states of Maranhão, Piauí, Ceará, Rio Grande do Norte and Paraíba at the beginning of the XXI century. The in situ characterization of G. hirsutum and G. barbadense was conducted through structured interviews with the cotton plants owners and through the analysis of the environment. The data were collected during expeditions undertaken between the years 2004 to 2005. Twenty-two plants were collected in the state of Paraíba, forty-four in the state of Rio Grande do Norte, one hundred and forty-six in the state of Ceará, forty in the state of Maranhão and ninety-one plants in the state of Piauí. All plants collected in the states of Paraíba and Rio Grande do Norte belonged to moco type. Moco cotton plants also predominated in the other states, representing 92%, 62% and 78% of plants collected in Ceará, Piauí and Maranhão, respectively. The other cotton plants collected belong to the species G. barbadense. The cotton plants were found in situ as dooryard plants, roads side, feral populations, cultivation or local varieties. Great part were dooryard plants (45.2%), being major in Piauí and Maranhão. Cultivation predominated in Ceará; in Rio Grande do Norte feral populations were the most frequent and, in Paraíba, local varieties. The maintenance of moco plants is related, mainly, to the phytotherapic domestic use (20.9%) and to confection of lamp wicks (29.7%). Few inhabitants in Paraíba, Rio Grande do Norte, Piauí and none in Maranhão used harvest the plants, storage the seeds or gin; however, in Ceará, 40.5% of owners affirmed that they harvested and commercialized the fiber. It was found that the maintenance of species is dependent of the fragile cultural habits of local inhabitants, therefore the maintenance in situ is not a suitable way to conservation of genetic resources. The efforts must be directed to the continuity of collections, maintenance and characterization ex situ

Análise comparativa dos genes de reparo do DNA em Gluconacetobacter diazotrophicus e Gluconobacter oxydans

Gluconacetobacter diazotrophicus é uma alfa-proteobactéria Gram-negativa, tolerante a meios ácidos, fixadora de nitrogênio atmosférico e foi a primeira bactéria diazotrófica endofítica isolada da cana-de-açúcar. Por sua vez, Gluconobacter oxydans, também alfa-proteobactéria Gram-negativa, possui a capacidade de oxidar incompletamente alcoóis e carboidratos. Ambas de interesse biotecnológico e industrial, essas bactérias tiveram seus genomas seqüenciados completamente em 2007. Desta forma, foi de interesse desse trabalho analisar e comparar os genes de reparo do DNA devido sua importância na manutenção da integridade genômica. Sendo assim, as vias de reparo presentes nos dois organismos foram identificadas, utilizando como base uma terceira alfa-proteobactéria, a Caulobacter crescentus, cujos genes de reparo foram descritos por um trabalho anterior e também os genes bem estabelecidos para o reparo do DNA em Escherichia coli. Para esse estudo, um banco de dados contendo ortólogos para os genes de reparo de DNA encontrados nos organismos foi criado e análises comparativas por similaridade usando o pacote Blast e o software Clustal foram feitas. Este estudo demonstrou que as principais vias de reparo ao DNA reparos por excisão, reparo direto, reparo recombinacional e reparo pelo sistema SOS estão presentes nos organismos analisados, demonstrando, na maioria das vezes, boa similaridade com E. coli. Interessantemente, foram encontradas duplicações gênicas nos quais uma das cópias estava presente no cromossomo e a outra, no plasmídeo, como no caso de UvrD, DnaE e Ssb, possivelmente caracterizando eventos de transferência lateral. Por fim, uma grande novidade foi a identificação de ortólogos para RecB em G. diazotrophicus e G. oxydans e de ortólogos duplicados de RecD em G. diazotrophicus. Até o momento, não havia sido relatada a presença de membros da via de iniciação RecBCD do reparo recombinacional em alfaproteobactérias