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【Objective】 HLA-DRB1 * 11:01, as a class HLA-Ⅱ gene, was reported to be associated with spontaneous clearance of HCV in Han and Li population. Our study was to investigate the effects of viral selection pressure and CD4+T cell epitope on the natural outcome of HCV infection in HLA-DRB1 * 11:01 positive infected patients. 【Methods】 The positive selection sites and population growth of E1E2 and NS3 genes of common HCV 6a in HLA-DRB1 * 11:01 positive and negative groups in Guangdong were respectively analyzed. The peptide library covering the conserved regions of common HCV genotypes was used to stimulate HCV spontaneous clearance group and chronic infection group using ELISPOT method. Reactive peptides were obtained according to the number of spot-forming cells per well and the frequency of occurrence in different groups. 【Results】 The positive selection sites (PSSs) of E1E2 and NS3 of common HCV 6a in HLA-DRB1 * 11:01 negative group were greater than those in HLA-DRB1 * 11:01 positive group. Furthermore, the number of PPSs in CD4+T cell peptide in HLA-DRB1 * 11:01 negative group were also greater than those in HLA-DRB1 * 11:01 positive group; Both groups of HCV 6a had a population growth in Guangdong, and the expansion trend of HLA-DRB1 * 11:01 negative group was significantly higher than that of HLA-DRB1 * 11 :01 positive group. Compared to HCV chronic infection group, the response rate of HCV spontaneous clearance group to five peptides (C-52 E2691-707, C-119 NS31545-1560, C-134 NS4A1669-1684, C-154 NS4B1912-1927, C-159 NS4B1929-1944) was higher. However, the HCV chronic infection group showed a higher response rate to two of the peptides(C-111 NS31497-1512, C-130 NS31650-1665). When HLA-DRB1 * 11:01 typing was considered, there was no significant difference in HCV-specific immune response generated by PBMCs between HLA-DRB1 * 11:01 positive and HLA-DRB1 * 11:01 negative groups. 【Conclusion】 This study revealed the relationship between viral selection pressure of HLA-DRB1 * 11:01 HCV infected persons and CD4+T cell antigen epitopes. At the same time, CD4+ T cell antigen epitopes of HCV pan-genotype were obtained, providing basic data for the development of T cell vaccine suitable for HCV pan-genotype.
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【Objective】 To learn the situation of the evolution process of HCV virus population and the selection pressure of HCV NS5B in intravenous drug users (IDUs) in Guangdong. 【Methods】 141 blood samples from hepatitis C virus (HCV) RNA-positive blood donors and 58 from HCV patients in Guangdong were randomly collected for HCV NS5B sequence amplification, combined with HCV NS5B sequences from blood donors and IDUs obtained by sequencing previously(between 2009 and 2011). Homology analysis was performed by Molecular Evolutionary Genetics Analysis (MEGA) software, evolutionary analysis were performed by Bayesian Evolutionary Analysis Sampling Trees (BEAST) software package. Selection pressure analysis was performed on sequences isolated from IDUs by Datamonkey online software package with Mixed Effects Model Evolution (MEME) method, and the population expansion of species were analyzed using Tajima and Fu neutrality test by Arlequin software. 【Results】 The comparison results of internal homology among different subtypes of IDUs in this group were as follows : HCV-3b had the highest homology (97%), followed by HCV-3a (96%), HCV-6a (95%) and HCV-1b (94%); HCV evolution rate analysis showed that HCV-1b had the fastest evolution rate [2.17E-03 substitutions/site/year (y/y/y)], followed by HCV-3b (2.12E-0 y/y/y), HCV-3a (1.58E-03 y/y/y) and HCV-6a (1.28E-03 y/y/y). The analysis on effective population of HCV: 1980~1990 was rapid growth period for HCV-6a, 1990~1995 period for HCV-1b, and 2000~2007 period for HCV-3a. HCV population genetic characteristics was as follows: HCV-1b, 3a, 3b and 6a experienced population expansion, among which 3a and 3b were the most obvious. As to the analysis of HCV selection pressure, two positive selection sites (235 and 243)were found in the 339 nucleotide fragment of the NS5B sequence in injecting drug users, but mutation only occurred at position 316 [mutation rate 1.24% (14/1 130)] among 5 direct antiviral drug (DAA) sites in this gene. 【Conclusion】 The evolution of HCV-3b in Guangdong has showed an obvious trend of population expansion, with a high proportion and homology especially in the local IDUs. HCV-3b should be the focus of HCV prevention and control in this region. Given that the positively selected sites of the HCV NS5B gene region of IDUs in Guangdong are non-DAA binding sites, DAA is expected to demonstrate a good effect on these patients.
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The hepatic lipase plays a central role in the lipid metabolism, catalyzing the hydrolysis of phospholipids, monoglycerides, diglycerides, and triglycerides, and acyl-CoA. It is also implied in the conversion of very low-density lipoprotein and intermediate density lipoprotein to low density lipoproteins. As a consequence, the gene encoding the hepatic lipase (LIPC) is associated with several diseases derived from the imbalance of lipids that are in general derived from the interaction between life styles and genetic architecture. Therefore, it is interesting to understand more about the characteristics of the microevolutionary processes affecting genes that, like LIPC, have a role in nutrition and lipid metabolism in human populations. We explored the selection signatures on LIPC in 26 populations, detecting three regions under recent positive selection
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El proteasoma es un complejo proteico grande, el cual se encuentra fundamentalmente en todas las células eucariotas ya que juega un rol muy importante en los procesos celulares, tales como: la diferenciación celular, la progresión del ciclo celular, el desarrollo y la apoptosis celular. Existen varios tipos de proteasomas como el constitutivo, los intermedios, el inmunoproteasoma y el timoproteasoma, los cuales están presenten en las células del cuerpo en dependencia de la estructura y función de ellas. Sin embargo, se encuentran en las células del sistema inmune donde no solo juegan un papel muy importante en el procesamiento antigénico para la respuesta inmune, sino en los mecanismos de tolerancia central durante el proceso de ontogenia de los linfocitos T en el timo. Así, las células epiteliales tímicas corticales son células presentadoras de antígenos, las cuales presentan características intrínsecas únicas al presentar el timoproteasoma, la catepsina L y la proteasa serin específica del timo. Además, se ha observado una alta tasa de macroautofagia en comparación a las otras células del cuerpo, por lo que serán esenciales en la obtención de un repertorio de linfocitos T CD4+ y CD8+ que tendrán la capacidad de discriminar lo propio y lo no propio. Por lo que se debería considerar que la tolerancia central no está únicamente definida por el mecanismo de selección negativa, sino que a su vez la selección positiva juega un papel muy importante en la definición del repertorio de clones de linfocitos T no autorreactivos. El objetivo es discutir acerca del proteasoma, los tipos de proteasomas y sus implicaciones en la tolerancia central de los linfocitos T(AU)
The proteasome is a large protein complex mainly located in all eukaryote cells, where it plays a very important role in cellular processes like differentiation, cycle progression, development and apoptosis. There are several types of proteasomes: constitutive, intermediate, immunoproteasome and thymoproteasome, which are present in cells depending on their structure and functions. However, they are found in cells of the immune system, where they play a very important role not only in antigenic processes related to the immune response, but also in central tolerance mechanisms during T lymphocyte ontogeny in the thymus. Thymic cortical epithelial cells are therefore antigen-presenting cells with unique intrinsic characteristics, since they contain thymoproteasome, cathepsin L, and thymus-specific serine protease. Additionally, a high rate of macroautophagy has been observed in comparison with bibr cells of the body. They are thus essential to obtain a repertoire of CD4+ and CD8+ T lymphocytes capable of distinguishing the body's own elements from alien structures. Therefore, central tolerance should not be viewed as only defined by the negative selection mechanism, since positive selection also plays a very important role in defining the repertoire of non-autoreactive T lymphocyte clones. The purpose of the study was to discuss the proteasome, the types of proteasomes and their involvement in T lymphocyte central tolerance(AU)
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Humansالملخص
Abstract Domestication is of unquestionable importance to the technological revolution that has given rise to modern human societies. In this study, we analyzed the DNA and protein sequences of six genes of the oxytocin and arginine vasopressin systems (OXT-OXTR; AVP-AVPR1a, AVPR1b and AVPR2) in 40 placental mammals. These systems play an important role in the control of physiology and behavior. According to our analyses, neutrality does not explain the pattern of molecular evolution found in some of these genes. We observed specific sites under positive selection in AVPR1b (ω = 1.429, p = 0.001) and AVPR2 (ω= 1.49, p = 0.001), suggesting that they could be involved in behavior and physiological changes, including those related to the domestication process. Furthermore, AVPR1a, which plays a role in social behavior, is under relaxed selective constraint in domesticated species. These results provide new insights into the nature of the domestication process and its impact on the OXT-AVP system.
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Abstract The DNA methyltransferase 2 (DNMT2) protein is the most conserved member of the DNA methyltransferase family. Nevertheless, its substrate specificity is still controversial and elusive. The genomic role and determinants of DNA methylation are poorly understood in invertebrates, and several mechanisms and associations are suggested. In Drosophila, the only known DNMT gene is Dnmt2. Here we present our findings from a wide search for Dnmt2 homologs in 68 species of Drosophilidae. We investigated its molecular evolution, and in our phylogenetic analyses the main clades of Drosophilidae species were recovered. We tested whether the Dnmt2 has evolved neutrally or under positive selection along the subgenera Drosophila and Sophophora and investigated positive selection in relation to several physicochemical properties. Despite of a major selective constraint on Dnmt2, we detected six sites under positive selection. Regarding the DNMT2 protein, 12 sites under positive-destabilizing selection were found, which suggests a selection that favors structural and functional shifts in the protein. The search for new potential protein partners with DNMT2 revealed 15 proteins with high evolutionary rate covariation (ERC), indicating a plurality of DNMT2 functions in different pathways. These events might represent signs of molecular adaptation, with molecular peculiarities arising from the diversity of evolutionary histories experienced by drosophilids.
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Abstract Alcohol dehydrogenases belong to the large superfamily of medium-chain dehydrogenases/reductases, which occur throughout the biological world and are involved with many important metabolic routes. We considered the phylogeny of 190 ADH sequences of animals, fungi, and plants. Non-class III Caenorhabditis elegans ADHs were seen closely related to tetrameric fungal ADHs. ADH3 forms a sister group to amphibian, reptilian, avian and mammalian non-class III ADHs. In fishes, two main forms are identified: ADH1 and ADH3, whereas in amphibians there is a new ADH form (ADH8). ADH2 is found in Mammalia and Aves, and they formed a monophyletic group. Additionally, mammalian ADH4 seems to result from an ADH1 duplication, while in Fungi, ADH formed clusters based on types and genera. The plant ADH isoforms constitute a basal clade in relation to ADHs from animals. We identified amino acid residues responsible for functional divergence between ADH types in fungi, mammals, and fishes. In mammals, these differences occur mainly between ADH1/ADH4 and ADH3/ADH5, whereas functional divergence occurred in fungi between ADH1/ADH5, ADH5/ADH4, and ADH5/ADH3. In fishes, the forms also seem to be functionally divergent. The ADH family expansion exemplifies a neofunctionalization process where reiterative duplication events are related to new activities.
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Objective To understand the genotypes of hepatitis C virus (HCV) circulating in intravenous drug users (IDUs) in Pudong new district,Shanghai,and explore the population growth and selection pressure of the HCV strains isolated.Methods A total of 200 serum specimens sampled from IDUs in local methadone maintenance treatment clinic in Pudong were used for amplification of a HCV NS5B 377-nt partial sequence.Mean evolutionary rate and effective number of infections were estimated based on the 377-nt partial sequences of the HCV strains isolated from IDUs and isolated contemporarily from local voluntary blood donors,men who have sex with men and reported hepatitis C cases by using BEAST software.Selection pressure sites were identified with online Datamonkey software for subsequent comparison with direct-acting antiviral (DAA) drug binding sites.Results A total of 39 (19.5％) serum specimens were positive for HCV RNA.The genotypes were determined based on the HCV NS5B 377-nt partial sequences as follows:subtype 3a (n=14),3b (n=13),lb (n=7),6a (n=4) and 6n (n=1).The partial sequences of the HCV strains isolated in IDUs shared high homology with the sequences of the HCV strains isolated in other populations.The Bayesian Skyline Plot indicated that the estimated infections with HCV subtype 1b increased exponentially during the 1990s,whereas that of subtypes 3a and 3b increased slowly since the mid-1990s.In the NS5B 377-nt partial sequences of the HCV strains isolated in IDUs,there were two positive selection sites and seventy-eight negative selection sites recognized.The mutation rate was as low as 2.2％ in the 377-nt partial sequences corresponding to the known seven DAA drug binding sites.Conclusions HCV subtype 3a and 3b were the predominant genotypes in the IDUs in Pudong.Subtype lb was prevalent in different populations and evolved very rapidly,and more infections might be caused,suggesting further attention to its prevention,control and treatment.Although DAA treatment based on HCV NS5B binding sites targeting local IDUs might be effective,it is still necessary to strengthen the surveillance.
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Objective To understand the genotypes of hepatitis C virus (HCV) circulating in intravenous drug users (IDUs) in Pudong new district,Shanghai,and explore the population growth and selection pressure of the HCV strains isolated.Methods A total of 200 serum specimens sampled from IDUs in local methadone maintenance treatment clinic in Pudong were used for amplification of a HCV NS5B 377-nt partial sequence.Mean evolutionary rate and effective number of infections were estimated based on the 377-nt partial sequences of the HCV strains isolated from IDUs and isolated contemporarily from local voluntary blood donors,men who have sex with men and reported hepatitis C cases by using BEAST software.Selection pressure sites were identified with online Datamonkey software for subsequent comparison with direct-acting antiviral (DAA) drug binding sites.Results A total of 39 (19.5％) serum specimens were positive for HCV RNA.The genotypes were determined based on the HCV NS5B 377-nt partial sequences as follows:subtype 3a (n=14),3b (n=13),lb (n=7),6a (n=4) and 6n (n=1).The partial sequences of the HCV strains isolated in IDUs shared high homology with the sequences of the HCV strains isolated in other populations.The Bayesian Skyline Plot indicated that the estimated infections with HCV subtype 1b increased exponentially during the 1990s,whereas that of subtypes 3a and 3b increased slowly since the mid-1990s.In the NS5B 377-nt partial sequences of the HCV strains isolated in IDUs,there were two positive selection sites and seventy-eight negative selection sites recognized.The mutation rate was as low as 2.2％ in the 377-nt partial sequences corresponding to the known seven DAA drug binding sites.Conclusions HCV subtype 3a and 3b were the predominant genotypes in the IDUs in Pudong.Subtype lb was prevalent in different populations and evolved very rapidly,and more infections might be caused,suggesting further attention to its prevention,control and treatment.Although DAA treatment based on HCV NS5B binding sites targeting local IDUs might be effective,it is still necessary to strengthen the surveillance.
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Objective To investigate the serotypes of human enterovirus B ( HEV-B) species cau-sing hand, foot and mouth disease ( HFMD) and to analyze the genetic characteristics of VP1 region in cox-sackievirus B2 ( CVB2 ) and coxsackievirus B5 ( CVB5 ) strains circulating in Anyang area during 2011 to 2015. Methods Real-time RT-PCR and semi-nested RT-PCR were performed to identify coxsackievirus A16 (CVA16), enterovirus 71 (EV71) and other serotypes of enterovirus in order to obtain the complete etiologic composition of HFMD. The numbers of HEV-B serotypes and the percentages of specimens positive for every serotype in all enterovirus-positive specimens were calculated. As CVB2 and CVB5 were the pre-dominant serotypes of HEV-B species, five pairs of primers targeting the VP1 regions of CVB2 and CVB5 were designed to obtain the complete nucleotide sequences of CVB2 and CVB5 VP1 regions. The phylogenet-ic trees were constructed based on the VP1 sequences obtained in this study and those submitted to GenBank by using MEGA7. 0 and BioEdit7. 2. The selection pressures on VP1 regions of CVB2 and CVB5 strains cir-culating in China in recent years were evaluated with the online program of DataMonkey. Results A total of 57 specimens that belonged to 14 serotypes of HEV-B species were detected in Anyang area from 2011 to 2015. The 14 serotypes of HEV-B species accounted for 56% of all serotypes of enterovirus and the speci-mens positive for HEV-B species accounted for 3. 06% of all enterovirus-positive specimens. The HFMD ca-ses caused by most of the HEV-B serotypes were sporadic cases. Small outbreaks of HFMD could also be caused by some serotypes of HEV-B such as CVB2 and CVB5. The complete sequences of VP1 region were obtained from 8 CVB2 strains and 9 CVB5 strains. The phylogenetic trees based on the VP1 sequences dem-onstrated that the CVB2 strains were classified into four genotypes ( A-D) . The mean evolutionary distances between different genotypes ranged from 0. 191 to 0. 208 and the similarities in nucleotide sequences ranged from 79. 7% to 85. 8%. The CVB5 strains were classified into 6 genotypes (A-F). The mean evolutionary distances and the similarities in nucleotide sequences between different genotypes of CVB5 strains ranged from 0. 170 to 0. 285 and 76. 0% to 86. 8%, respectively. Strains of different genotypes varied significantly in the residues on positons 157 and 263 in the VP1 region of CVB2 strains and on positions 75, 90 and 95 in the VP1 region of CVB5 strains. All of the CVB2 strains isolated in Anyang area belonged to D genotype and located intensively in one lineage. The CVB5 strains circulated in Anyang area belonged to F genotype and located in two lineages. The selection pressures on CVB2 strains of D genotype and CVB5 strains of F geno-type circulating in China in recent years were 0. 037 and 0. 036, respectively. Six positively selected amino acid sites were found in the VP1 region of CVB5 strains, but no positively selected amino acid site was found in the VP1 region of CVB2 strains. Conclusion HEV-B species was an essential component of the etiologic spectrum of HFMD in Anyang area during 2011 to 2015, of which CVB5 and CVB2 were the predominant se-rotypes. The VP1 region of CVB5 was more complex and active than that of CVB2 over the course of evolution.
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Matrix metalloproteinases-9 (MMP-9) is an important cancer-associated, zinc-dependent endopeptidase. To investigate the natural selection hypothesis of MMP-9, the orthologous sequences from 12 vertebrates were compared and a molecular evolution analysis was performed. Results suggest that amino acid residues present in the middle region of the protein are more selectively constrained, whereas amino acid residues in the C-terminal region of the MMP-9 protein including exon 13 showed lowest conservation level in non-primate species, suggesting that it is an exon with fast evolving rate compared to the others analyzed. InterProScan analysis shows that exon 13 was located in hemopexin (PEX) domain of MMP-9. Positive selection was detected in PEX domain of MMP-9 protein between human and other species, which indicates that selective pressure may play a role in shaping the function of MMP-9 in the course of evolution.
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Non structural protein 4 (NSP4) gene of Rotavirus encodes a multifunctional protein which has significant role in viral multiplication and pathogenesis of acute watery diarrhoea associated with rotaviral gastroenteritis. It is known as the first viral enterotoxin and mutations of the gene have been linked to altered pathogenesis. This study was planned to ascertain the genotypes and genetic variations of NSP4 gene in the rotavirus strains prevalent in this area. We collected consecutive diarrhoeal stools from equal no of children aged under five years hospitalized with diarrhoea in a period from January 2010 to June 2012 and tested them for rotavirus antigen by ELISA. NSP4 gene was amplified by RT-PCR and subsequently sequenced (Big-Dye terminator kit using 3130 ABI, Genetic analyzer) and genotyped by Rotavirus C software. Of the 260 samples, 58(22.3%) samples were positive by ELISA. We were able to amplify NSP4 gene by RTPCR from 45 strains of which 35 amplicons were selected for sequencing. Total 25(71.4%) strains belonged to genotype E1, 6 (17.1%) strains to genotype E2 and 4 (11.4%) matched with the genotype E6. Sequence analysis revealed changes in the nucleotides causing punctate mutations in the conserved regions, the Inter species variable domain (ISVD) and the enterotoxin region (amino acid 114-135). On evolutionary analysis of 33 strains amino acid at position 131 was found under positive selection.
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Selective sweep can cause genetic differentiation across populations, which allows for the identification of possible causative regions/genes underlying important traits. The pig has experienced a long history of allele frequency changes through artificial selection in the domestication process. We obtained an average of 329,482,871 sequence reads for 24 pigs from three pig breeds: Yorkshire (n = 5), Landrace (n = 13), and Duroc (n = 6). An average read depth of 11.7 was obtained using whole-genome resequencing on an Illumina HiSeq2000 platform. In this study, cross-population extended haplotype homozygosity and cross-population composite likelihood ratio tests were implemented to detect genes experiencing positive selection for the genome-wide resequencing data generated from three commercial pig breeds. In our results, 26, 7, and 14 genes from Yorkshire, Landrace, and Duroc, respectively were detected by two kinds of statistical tests. Significant evidence for positive selection was identified on genes ST6GALNAC2 and EPHX1 in Yorkshire, PARK2 in Landrace, and BMP6, SLA-DQA1, and PRKG1 in Duroc.These genes are reportedly relevant to lactation, reproduction, meat quality, and growth traits. To understand how these single nucleotide polymorphisms (SNPs) related positive selection affect protein function, we analyzed the effect of non-synonymous SNPs. Three SNPs (rs324509622, rs80931851, and rs80937718) in the SLA-DQA1 gene were significant in the enrichment tests, indicating strong evidence for positive selection in Duroc. Our analyses identified genes under positive selection for lactation, reproduction, and meat-quality and growth traits in Yorkshire, Landrace, and Duroc, respectively.
الموضوعات
Female , Gene Frequency , Haplotypes , Lactation , Meat , Polymorphism, Single Nucleotide , Reproduction , Swine , Natural Resourcesالملخص
As a secreted glycoprotein that binds to the extracellular domain of Toll-like receptor 4 (TLR4), Lymphocyte Antigen 96 (LY96), also called myeloid differentiation 2 (MD2), is required for the activation of TLR4 by lipopolysaccharide (LPS) and plays an important role in innate immunity, which is the first line of defence against microbial infections. Previous studies have proposed that mammalian toll-like receptors (TLRs) have evolved under diversifying selection due to their role in pathogen detection. Given the fact that LY96 is highly functionally linked to TLR4, it would be interesting to test whether LY96 is under the intense pressure of natural selection. To investigate the natural selection hypothesis, we compared the coding sequences from 13 vertebrates and evaluated the molecular evolution of LY96 gene in these species. Result shows that natural selection at exon 4 has indeed played a role in shaping the function of LY96 in the course of evolution. In addition to the study of Nakajima, we found the two branch nodes with Ka/Ks ratios greater than 1: the one leading to cow and pig and the other to rabbit and the primates.
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El timo es un órgano linfoide central o primario localizado en la parte anterosuperior del tórax. Constituye el principal sitio de diferenciación y maduración de las células T. En esta revisión se detallan aspectos actuales de la embriología, la histología y la función tímica y en la generación de los diferentes subtipos de timocitos y su diferenciación a células T maduras efectoras, la inducción de las células T tímicas reguladoras involucradas en el mantenimiento de la tolerancia a lo propio y la involución que sufre este órgano durante el proceso de inmunosenescencia
Thymus is a primary organ located in the antesuperior area of the torax. It is the principal place of differentiation and maduration of T cells. In this review present aspects of the embriology, histology and thymic function are detailed, as well as its role in the generation of different kinds of thymic cells; its differentiation to mature cells and of regulator T cells has a crucial role in tolerance induction. Moreover, thymic involution during of immunosenescence process is shown
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Humans , Organogenesis/physiology , DiGeorge Syndrome/history , Thymus Gland/physiopathology , Agingالملخص
Biological nitrogen fixation is accomplished by prokaryotes through the catalytic action of complex metalloenzyme, nitrogenase. Nitrogenase is a two-protein component system comprising MoFe protein (NifD&K) and Fe protein (NifH). NifH shares structural and mechanistic similarities as well as evolutionary relationships with light-independent protochlorophyllide reductase (BchL), a photosynthesis-related metalloenzyme belonging to the same protein family. We performed a comprehensive bioinformatics analysis of the NifH/BchL family in order to elucidate the intrinsic functional diversity and the underlying evolutionary mechanism among the members. To analyse functional divergence in the NifH/ BchL family, we have conducted pair-wise estimation in altered evolutionary rates between the member proteins. We identified a number of vital amino acid sites which contribute to predicted functional diversity.We have also made use of the maximum likelihood tests for detection of positive selection at the amino acid level followed by the structure-based phylogenetic approach to draw conclusion on the ancient lineage and novel characterization of the NifH/BchL protein family. Our investigation provides ample support to the fact that NifH protein and BchL share robust structural similarities and have probably deviated from a common ancestor followed by divergence in functional properties possibly due to gene duplication.
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Tetrodotoxin (TTX) is a highly potent neurotoxin that blocks the action potential by selectively binding to voltage-gated sodium channels (Na v). The skeletal muscle Na v (Na v1.4) channels in most pufferfish species and certain North American garter snakes are resistant to TTX, whereas in most mammals they are TTX-sensitive. It still remains unclear as to whether the difference in this sensitivity among the various vertebrate species can be associated with adaptive evolution. In this study, we investigated the adaptive evolution of the vertebrate Na v1.4 channels. By means of the CODEML program of the PAML 4.3 package, the lineages of both garter snakes and pufferfishes were denoted to be under positive selection. The positively selected sites identified in the p-loop regions indicated their involvement in Na v1.4 channel sensitivity to TTX. Most of these sites were located in the intracellular regions of the Na v1.4 channel, thereby implying the possible association of these regions with the regulation of voltage-sensor movement.
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A detailed study of putative recombination events and their evolution frequency in the whole genome of the currently known members of the family Tombusviridae, comprising 79 accessions retrieved from the international databases, was carried out by using the RECCO and RDP version 3.31β algorithms. The first program allowed the detection of potential recombination sites in seven out of eight virus genera (Aureusvirus, Avenavirus, Carmovirus, Dianthovirus, Necrovirus, Panicovirus, and Tombusvirus), the second program provided the same results except for genus Dianthovirus. On the other hand, both methods failed to detect recombination breakpoints in the genome of members of genus Machlomovirus. Furthermore, based on Fisher's Exact Test of Neutrality, positive selection exerted on protein-coding genes was detected in 17 accession pairs involving 15 different lineages. Except genera Machlomovirus, and Panicovirus along with unclassified Tombusviridae, all the other taxonomical genera and the unassigned Tombusviridae encompassed representatives under positive selection. The evolutionary history of all members of the Tombusviridae family showed that they segregated into eight distinct groups corresponding to the eight genera which constitute this family. The inferred phylogeny reshuffled the classification currently adopted by the International Committee on Taxonomy of Viruses. A reclassification was proposed.
الموضوعات
Base Sequence , Computational Biology , Phylogeny , Recombination, Genetic , Tombusviridaeالملخص
This article reports the genetic divergence of PCV2 after passages in VERO and SK-RST cell lines. Fishers exact test indicated a trend for positive selection on the cap gene. These results allow insights on the development of PCV2 vaccines and the evolution of the genus Circovirus.
Este artigo relata a divergência genética de PCV2 após passagens em células das linhagens VERO e SK-RST. O teste exato de Fisher indicou tendência para seleção positiva no gene cap. Estes resultados permitem inferências relativas ao desenvolvimento de vacinas contra PCV2 e sobre a evolução do gênero Circovirus.
الموضوعات
Animals , Circovirus/genetics , Selection, Genetic , Biological Evolution , Polymerase Chain Reactionالملخص
Se estableció un sistema de organogénesis indirecta para la obtención de brotes múltiples a partir de segmentos internodales de la variedad Diacol Capiro. La ubicación de explantes en medio Murashige y Skoog (MS) suplementado con 2 mg/l de zeatina ribosido (ZR), 0,02 mg/l de ácido naftalenácetico (ANA) y 0,02 mg/l de ácido giberélico (AG3), permite la obtención de plántulas entre la séptima y novena semana con una efectividad del 80-100%. Mediante ubicación de explantes previamente cocultivados con la cepa LBA4404 de Agrobacterium tumefaciens que contiene el plásmido recombinante pNOV022, se verificó la utilidad del medio para procesos de transformación, obteniéndose tasas hasta del 100% de regeneración. Finalmente, con el objetivo de determinar el uso potencial de la manosa como agente selectivo en procesos de transformación, se evaluó el efecto de diferentes concentraciones de manosa sobre la viabilidad y capacidad regenerativa de explantes.
A system of indirect organogenesis for the multiple buds production from internode stem sections in Diacol Capiro variety was established. Explants on Murashige & Skoog (MS) medium with zeatine riboside (ZR) 2 mg/l, naftalenacetic acid (NAA) 0.02 mg/l and giberelic acid (GA3) 0.02 mg/l, produced plants ranging between 7 to 9 weeks with 80-100% effectiveness. In the same medium, explants infected with Agrobacterium LBA4404 strain which carries recombinant plasmid pNOV022, produced regeneration rates reached 100%, thus, the medium utility for trnsformation processes was verified. Finally, to determine the potential use of the mannose as selective agent in transformation processes, the effect of different mannose concentrations on explant viability and regenerative capacity was evaluated.