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Registros recuperados: 66
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16S rRNA region based PCR protocol for identification and subtyping of Parvimonas micra BJM
Ota-Tsuzuki,C.; Brunheira,A.T.P.; Mayer,M.P.A..
The present study established a PCR protocol in order to identify Parvimonas micra and to evaluate the intra-species diversity by PCR-RFLP of 16S rRNA partial sequence. The data indicated that the protocol was able to identify this species which could be clustered in five genotypes.
Tipo: Info:eu-repo/semantics/article Palavras-chave: PCR-RFLP; 16S rRNA; Anaerobic cocci.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822008000400001
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A C1069G SNP of the MC4R gene and its association with economic traits in Korean native cattle (brown, brindle, and black) Electron. J. Biotechnol.
Lee,Youngsub; Park,Sairom; Kim,Hun; Lee,Seung Kyu; Kim,Jin Woo; Lee,Hak Kyo; Jeong,Dong Kee; Lee,Sung Jin.
Background: The melanocortin-4 receptor gene (MC4R) plays an important role in regulating food intake and body weight in mammals. In the present study, we identified the MC4R gene in native Korean brown, brindle, and black cattle by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), and investigated its association with economic traits. A total of 413 cattle from the three breeds were tested for backfat thickness, carcass weight, longissimus muscle area, and marbling score, and statistical data were analyzed using the SAS program. Results: The C allele had the highest frequency in brown and brindle cattle, and the G allele frequency was highest in black cattle. The C1069G SNP was significantly (p < 0.05) associated with only...
Tipo: Journal article Palavras-chave: Economic trait; Marbling scores; MC4R gene; PCR-RFLP; SNP.
Ano: 2013 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000500014
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A modified simple RFLP-PCR method for single nucleotide polymorphism (SNP) typing Genet. Mol. Biol.
Xiao,Junhua; Xin,Xiujuan; Luan,Xiaohui; Dongzhi,Wei; Shengli,Yang.
We describe a modified single nucleotide polymorphism (SNP) typing method based on the restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). This is a simple, economical method without the need for special equipment. For most SNP loci, a common restriction endonuclease (Hind III, EcoR I or BamH I) recognizing site (RER) can be introduced into one allelic form, but not the other by two rounds of mismatched PCR. The flanking regions can be changed by as many as five bases after PCR amplification with specially designed mismatching primers so the genotypes can be distinguished after digestion of the PCR products with corresponding endonucleases.
Tipo: Info:eu-repo/semantics/article Palavras-chave: SNP genotyping; PCR-RFLP; RER site.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572006000300028
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Alternative genotyping method for the single nucleotide polymorphism A2959G (AF159246) of the bovine CAST gene PAB
Curi,Rogério Abdallah; Chardulo,Luis Artur Loyola; Silveira,Antônio Carlos; Oliveira,Henrique Nunes de.
The objective of this work was to genotype the single nucleotide polymorphism (SNP) A2959G (AF159246) of bovine CAST gene by PCR-RFLP technique, and to report its use for the first time. For this, 147 Bos indicus and Bos taurus x Bos indicus animals were genotyped. The accuracy of the method was confirmed through the direct sequencing of PCR products of nine individuals. The lowest frequency of the meat tenderness favorable allele (A) in Bos indicus was confirmed. The use of PCR-RFLP for the genotyping of the bovine CAST gene SNP was shown to be robust and inexpensive, which will greatly facilitate its analysis by laboratories with basic structure.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Calpastatin gene; Meat texture; PCR-RFLP; SNP.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2008000500014
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Alternative methodologies for genotyping polymorphisms in the CAST and CAPN1 genes in beef cattle R. Bras. Zootec.
Blecha,Isabella Maiumi Zaidan; Sousa,Isadora Inácio; Ferreira,Anna Beatriz Robottom; Feijó,Gelson Luis Dias; Torres Júnior,Roberto Augusto de Almeida; Siqueira,Fabiane.
ABSTRACT The objectives of this study were to genotype single nucleotide polymorphisms (SNP) AF159246:g.2959A>G (CAST/DdeI) and AF248054.2:g.6545C>T (CAPN4751) in beef cattle by PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism), using the restriction enzyme DdeI for both SNP, and describe the use of these genotyping methodologies for the first time. For the SNP located in the CAST gene, new primers were designed, and for the SNP of the CAPN1 gene, the same primers previously described in the literature were used. Bonsmara, Caracu, Senepol, Nelore, and Angus bulls were chosen from among the most used bulls in breeding programs according to their genealogy and the lowest possible degree of parentage between them to ensure...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Animal breeding; Marker-assisted selection; PCR-RFLP; SNP; Tenderness.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-35982019000100311
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Aromatase gene and its effects on growth, reproductive and maternal ability traits in a multibreed sheep population from Brazil Genet. Mol. Biol.
Lôbo,Ana Maria Bezerra Oliveira; Lôbo,Raimundo Nonato Braga; Paiva,Samuel Rezende.
We determined the polymorphism C242T of the aromatase gene (Cyp19) and its allelic frequency, as well as the effect of the variants on productive and reproductive traits in 71 purebred Santa Inês sheep, 13 purebred Brazilian Somali sheep, nine purebred Poll Dorset sheep, and 18 crossbred 1/2 Dorper sheep. The animals were genotyped using the PCR-RFLP technique. The influence of the animal's genotype on its performance or on the performance of its lambs was analyzed by the least square method. Another factor assessed was the importance of the animal's genotype in analysis models for quantitative breeding value estimates, and whether there were differences among the averages of breeding values of animals with different genotypes for this gene. In the sample...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Body weight; Lambing interval; PCR-RFLP; Polymorphism; SNP; Litter weight.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572009000300010
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Associação do loco BoLA-DRB3.2 com produção de leite em bovinos da raça Gir Arq. Bras. Med. Vet. Zootec.
Machado,M.A.; Nascimento,C.S.; Martinez,M.L.; Silva,M.V.G.B.; Campos,A.L.; Teodoro,R.L.; Verneque,R.S.; Guimarães,S.E.F..
A associação entre os alelos do loco BoLA-DRB3.2, identificados pela técnica de PCR-RFLP, e a produção de leite na raça Gir foi estudada por meio da análise de dados moleculares e fenotípicos de 424 vacas Gir, utilizando um modelo misto, sob modelo animal. Os dados moleculares consistiam dos genótipos dos animais para os alelos do loco BoLA-DRB3.2 e os dados fenotípicos eram referentes à produção de leite em até 305 dias de lactação. O loco é altamente polimórfico nesta raça, sendo identificados sete alelos (BoLA-DRB3.2*4, *8, *11, *19, *28, *41 e *48) que não haviam sido encontrados em animais zebuínos. Dois alelos (*16 e *29) estavam significativamente associados com maiores produções de leite, sugerindo que o próprio loco BoLA-DRB3.2 ou um QTL a ele...
Tipo: Info:eu-repo/semantics/article Palavras-chave: BoLA-DRB3; PCR-RFLP; Gene candidato; Bovinos; Gir.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352005000300017
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Association of polymorphisms in the heparanase gene (HPSE) with hepatocellular carcinoma in Chinese populations Genet. Mol. Biol.
Yu,Lixia; Zhang,Xiaoai; Zhai,Yun; Zhang,Hongxing; Yue,Wei; Zhang,Xiumei; Wang,Zhifu; Zhou,Hong; Zhou,Gangqiao; Gong,Feng.
Abstract Heparanase activity is involved in cancer growth and development in humans and single nucleotide polymorphisms (SNPs) in the heparanase gene (HPSE) have been shown to be associated with tumors. In this study, we investigated whether SNPs in HPSE were a risk factor for hepatocellular carcinoma (HCC) by undertaking a comprehensive haplotype-tagging, case-control study. For this, six haplotype-tagging SNPs (htSNPs) in HPSE were genotyped in 400 HCC patients and 480 controls by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. A log-additive model revealed significant correlations between the HPSE polymorphisms rs12331678 and rs12503843 and the risk of HCC in the overall samples (p = 0.0046 and p = 0.0055). When...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Genetic association; Heparanase; Hepatocellular carcinoma; HPSE gene; PCR-RFLP.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572017000500743
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Association of the bovine major histocompatibility complex (BoLA) BoLA-DRB3 gene with fat and protein production and somatic cell score in Brazilian Gyr dairy cattle (Bos indicus) Genet. Mol. Biol.
Nascimento,Carlos Souza do; Machado,Marco Antonio; Martinez,Mario Luiz; Silva,Marcos Vinícius G. Barbosa da; Guimarães,Marta Fonseca Martins; Campos,Ana Lúcia; Azevedo,Ana Luisa Sousa; Teodoro,Roberto Luiz; Verneque,Rui da Silva; Guimarães,Simone Eliza Facioni; Oliveira,Denise Aparecida Andrade.
The effect of the bovine major histocompatibility complex (BoLA) locus on animal health may be due to a direct action of its alleles on immune functions, whereas its indirect effect on production traits might be explained by the better general health conditions of more productive animals. In the present study, the BoLA-DRB3 gene was investigated in 1058 cows belonging to seven Brazilian Gyr Dairy herds (Bos indicus, Zebu cattle). A total of 37 alleles were identified, 15 of them described for the first time in a Zebu breed. A highly significant association (p < 0.02) was observed between allele *54 and a decrease (-26.1 kg) in milk protein yield and there was a significant association (p < 0.05) between this allele and lower (-26.07 kg) milk fat...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cattle; Marker-assisted selection; Molecular markers; PCR-RFLP.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572006000400011
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Authenticating the origin of different shrimp products on the Tunisian markets by PCR/RFLP method ArchiMer
Besbes, Nadia; Jerome, Marc; Berge, Jean-pascal; Sadok, Saloua.
This study describes a polymerase chain reaction using restriction fragment length polymorphism (PCR-RFLP) assay based on the 16S rRNA mitochondrial gene to identify commercial food products of wide range of Penaeidae and Pandalidae shrimp species commercialised in the Tunisian market. Phylogenetic analyses on 16S rRNA mitochondrial gene were used to study the relationships among the considered species. Penaeidae shrimp species was easily differentiated and confirmed by direct sequencing, showing a genetic distance of 0.34 with respect to Pandalidae species. A rapid and reliable PCR method using restriction fragment length polymorphism (RFLP) with three restriction enzymes (HpyCH4III/ MboI / AluI) was optimized for unambiguous differentiation of shrimp...
Tipo: Text Palavras-chave: Shrimps; Species identification; 16S rRNA mtDNA; PCR-RFLP.
Ano: 2015 URL: https://archimer.ifremer.fr/doc/00303/41445/40637.pdf
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Bovine kappa-casein gene polymorphism and its association with milk production traits Genet. Mol. Biol.
Rachagani,Satyanarayana; Gupta,Ishwar Dayal.
Point mutations in exon IV of the bovine κ-casein (CSN3) gene determine two allelic variants, A and B. These variants were distinguished by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis in the indigenous Sahiwal and Tharparkar cattle breeds. DNA samples (252 Sahiwal and 56 Tharparkar) were analyzed for allelic variants of the CSN3 gene. Polymorphism was detected by digestion of PCR-amplified products with HindIII, HhaI and HaeIII restriction enzymes, followed by separation on 3% agarose gels, and resolved by ethidium bromide staining. Allele A of the κ-casein gene occurred at a higher frequency than allele B, in both Sahiwal and Tharparkar breeds. The genotypic frequencies of AA, AB, and BB in the Sahiwal and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Indian cattle; Κ-casein; Genetic polymorphism; PCR-RFLP.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000500015
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Caracterização da região espaçadora 16-23S rDNA para diferenciação de estirpes de rizóbios utilizadas na produção de inoculantes comerciais no Brasil Ciência Rural
Oliveira,Andréia Mara Rotta; Bangel,Eliane Villamil; Hungria,Mariangela; Silveira,José Ricardo Pfeifer; Vargas,Luciano Kayser; Lisboa,Bruno Brito.
A identificação de estirpes de rizóbio tem sido feita pela especificidade por hospedeiros e ensaios microbiológicos tradicionais. Por constituírem um grupo filogeneticamente heterogêneo, diferentes técnicas moleculares têm sido empregadas para auxiliar na caracterização genética e na identificação de estirpes eficientes e competitivas para a produção de inoculantes. Este trabalho teve por objetivos caracterizar a região espaçadora 16S-23S rDNA das estirpes de rizóbios utilizadas nos inoculantes comercializados no Brasil para espécies leguminosas, utilizando a técnica da PCR em combinação com a de RFLP, e avaliar a possibilidade do uso desse marcador molecular como método auxiliar para identificação das estipes. A amplificação da região espaçadora 16-23 S...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Qualidade de inoculantes; PCR-RFLP; Marcadores moleculares.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782012000800015
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Caracterização de cepas de Corynebacterium pseudotuberculosis ovis isoladas de ovinos e caprinos MV&Z
Melo, Patrícia Rodrigues de; Carvalho, Cleber Eduardo Galvão; Andrade, Gisele Braziliano de; Soares, Cleber Oliveira; Rosinha, Grácia Maria Soares.
Foi efetuada a caracterização bioquímica, genotípica e a análise de virulência e patogenicidade de amostras isoladas de C. pseudotuberculosis. Foram utilizadas cinco amostras: CBO (Corynebacterium Ovino) 2512, CBO 28033, CBC (Corynebacterium Caprinos) 70D, CBC 118, CBC NPC e as amostras controle 1002 (Linfovac) e 00512 (Fiocruz). Para a identificação bioquímica, foi utilizado o kit APIcoryne (Bio Merieux-França). A caracterização genotípica foi realizada com o teste de PCR-RFLP para o gene rpoB, tratado com as enzimas MseI e StuI. As amostras foram avaliadas quanto à virulência e patogenicidade, em camundongos BALB/c inoculados com 1x103, 1x104, 1x105 e 1x106 Unidades Formadoras de Colônias por mL (UFC/mL). A caracterização bioquímica confirmou que as...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Linfadenite caseosa; Virulência; Patogenicidade; PCR-RFLP; Corynebacterium pseudotuberculosis.
Ano: 2014 URL: http://www.revistamvez-crmvsp.com.br/index.php/recmvz/article/view/23741
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Characterization of beta-thalassemia mutations in patients from the state of Rio Grande do Norte, Brazil Genet. Mol. Biol.
Silveira,Zama Messala Luna da; Barbosa,Maria das Vitórias; Fernandes,Thales Allyrio Araújo de Medeiros; Kimura,Elza Miyuki; Costa,Fernando Ferreira; Sonati,Maria de Fátima; Rebecchi,Ivanise Marina Moretti; Medeiros,Tereza Maria Dantas de.
35 unrelated individuals were studied for characterization as either heterozygous or homozygous for beta-thalassemia. Molecular analysis was done by PCR/RFLP to detect the mutations most commonly associated with beta-thalassemia (β0IVS-I-1, β+IVS-I-6, and β039). In the patients who showed none of these mutations, the beta-globin genes were sequenced. Of the 31 heterozygous patients, 13 (41.9%) had the β+IVS-I-6 mutation, 15 (48.4%) the β0IVS-I-1 mutation, 2 (6.5%) the β+IVS-I-110 mutation and 1 (3.2%) the β+IVS-I-5 mutation. IVS-I-6 was detected in the four homozygotes. The mutation in codon 39, often found in previous studies in Brazil, was not detected in the present case. This is the first study aiming at identifying mutations that determine...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Hereditary hemoglobinopathies; Beta-thalassemia; Mutations; PCR-RFLP; Brazilian population.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000300010
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Characterization of cry2-type genes of Bacillus thuringiensis strains from soil-isolated of Sichuan basin, China BJM
Liang,Hongxia; Liu,Yao; Zhu,Jun; Guan,Peng; Li,Shuangcheng; Wang,Shiquan; Zheng,Aiping; Liu,Huainian; Li,Ping.
Sichuan basin, situated in the west of China, is the fourth biggest basin in China. In order to describe a systematic study of the cry2-type genes resources from Bacillus thuringiensis strains of Sichuan basin, a total of 791 Bacillus thuringiensis strains have been screened from 2650 soil samples in different ecological regions. The method of PCR-restriction fragment length polymorphism (PCR-RFLP) was used to identify the type of cry2 genes. The results showed that 322 Bacillus thuringiensis strains harbored cry2-type genes and four different RFLP patterns were found. The combination of cry2Aa/cry2Ab genes was the most frequent (90.4%), followed by cry2Aa (6.8%) and cry2Ab alone (2.5%), and only one novel type of cry2 gene was cloned from one isolate...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bacillus thuringiensis; PCR-RFLP; SDS-PAGE; Novel cry2-type gene.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000100018
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Cryptosporidium infection in ostriches (Struthio camelus) in Brazil: clinical, morphological and molecular studies Rev. Bras. Ciênc. Avic.
Santos,MMAB; Peiró,JR; Meireles,MV.
Avian cryptosporidiosis has been reported in more than 30 species of birds. To date, the species infecting birds are C. baileyi, C. galli and C. meleagridis. In this study, the morphological, clinical and molecular characteristics of a Brazilian ostrich isolate of Cryptosporidium are described. The oocysts of this Brazilian isolate are larger and more elongated than those of Cryptosporidium previously reported in ostriches, which were morphologically similar to C. meleagridis. Morphological, biological and molecular analyses demonstrated similarity of this ostrich isolate with C. baileyi, suggesting that there are at least two Cryptosporidium species infecting ostriches; one with molecular, biological and morphological characteristics related to C....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Clinical signs; Cryptosporidium; Morphology; Ostriches; PCR-RFLP.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2005000200008
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Cytogenetic and molecular analyses in troglobitic and epigean species of Pimelodella (Siluriformes: Heptapteridae) from Brazil Neotropical Ichthyology
Dazzani,Bianca; Garcia,Caroline; Peixoto,Marilena; Trajano,Eleonora; Almeida-Toledo,Lurdes Foresti de.
Samples from seven different locations of the genus Pimelodella were genetically examined, two caves (exclusively subterranean, upper Tocantins River and São Francisco River) and five epigean (from upper Paraná River basin). Cytogenetic analyses revealed the same diploid number (2n=46) for all species besides similarities in both number and location of nucleolar organizer regions and C bands. FISH with 5S rDNA probes and CMA3 staining indicated significant differences among the studied species. Application of PCR-RFLP in ATPase 6 and 8 mitochondrial genes allowed building a minimum evolution phenogram identifying the close evolutionary relationship among groups. Both chromosomal and molecular data were useful to infer the relationships among studied...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cave fish; Cytogenetics; Evolutionary relationships; FISH; PCR-RFLP.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1679-62252012000300016
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Detección de variantes alélicas de la kappa-caseína en bovinos Hartón del Valle Acta Agron. (Palmira)
Naranjo,,Julián; Posso,Andrés; Cárdenas,Heiber; Muñoz F,Jaime E..
RESUMEN Se utilizó ADN de 2 machos y 34 hembras de la raza de ganado criollo Hartón del Valle para detectar polimorfismos en el gen que codifica para la kappa-caseína (κ-CN) mediante PCR-RFLP (reacción en cadena de la polimerasa-polimorfismo en la longitud de los fragmentos de restricción) y PCR-SSCP (reacción en cadena de la polimerasa-polimorfismo en la conformacion de ADN de cadena única). Se identificaron 17 individuos homocigotos AA, 15 heterocigotos AB y 4 homocigotos BB. La identificación de machos jóvenes portadores del genotipo de interés en menos de ocho horas mediante SSCP representa una ventaja para los propietarios de hatos abriendo la posibilidad de aumentar la frecuencia del alelo B, una de las vías para mejorar la eficiencia lechera para la...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bos taurus; Kappa-caseína; PCR-SSCP; PCR-RFLP.
Ano: 2007 URL: http://www.scielo.org.co/scielo.php?script=sci_arttext&pid=S0120-28122007000100043
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Detection of Metarhizium anisopliae var. anisopliae within infected sugarcane borer Diatraea saccharalis (Lepidoptera, Pyralidae) using specific primers Genet. Mol. Biol.
Destéfano,Ricardo Henri Rodrigues; Destéfano,Suzete A. Lanza; Messias,Claudio Luiz.
In order to construct specific primers for the detection and identification of the entomopathogenic fungus Metarhizium within infected sugarcane borer (Diatraea saccharalis) larvae we analyzed the ITS1 -5.8S- ITS2 rDNA regions of strains and varieties of M. anisopliae, M. album and M. flavoviride. The PCR amplification of these regions yielded a unique fragment of approximately 540 bp for M. anisopliae variety anisopliae strains E9, B/Vi and C (isolated in Brazil), 600 pb for M. a. anisopliae strain 14 (isolated in Australia), 650 bp for the M. album and 600 bp for M. flavoviride strains. The PCR products were digested with different restriction endonucleases (Afa I, Alu I, Dde I, Hae III, Hpa II and Sau 3A) and the PCR-RFLP profiles showed clear...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Metarhizium anisopliae; Entomopathogenic fungi; PCR-RFLP; ITS region.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572004000200020
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Determinación de las frecuencias alélicas de tres lactoproteínas en bovinos Criollo Limonero y Carora de Venezuela Rev Salud Anim.
Morillo,Milangela; Acosta,Atzel; Uffo,Odalys.
La caracterización genética del ganado autóctono supone conocer la situación de las poblaciones para su conservación. Aunque en Venezuela se han realizado estudios de caracterización de ganado Carora y Criollo Limonero, estos han sido aislados y no se han efectuado con la intención de esclarecer si la estructura genética para los genes de las tres principales proteínas lácteas, en dichas razas bovinas venezolanas, Criollo Limonero y Carora, refleja su variabilidad genética. Por dicha razón se determinaron los genotipos para las tres principales proteínas lácteas: k-caseina, a-lactoalbúmina y b-lactoglobulina en rebaños controlados. Para la caracterización de dichas razas se calcularon las frecuencias alélicas, las heterocigosidades por locus para cada...
Tipo: Journal article Palavras-chave: Bovino; Carora; Criollo Limonero; Proteínas lácteas; PCR-RFLP.
Ano: 2014 URL: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S0253-570X2014000300007
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