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  • 1
    Publication Date: 2021-05-19
    Description: In this study lusiferse gene extracted from Vibrio fischeri. V. fischeri strain was kindly provided by Iranian Research Organization for Science and Technology (IROST). Genomic DNA extraction was carried out by Phenol-chloroform. A DNA fragment encoding the luxA and LuxB was amplified by PCR using sense and antisense primers, specific restriction sites for BamH1 and Kpn1 were introduced into 5´ end of forward and reverse primer, respectively. The PCR product was purified from agarose gel and ligated into cleaved PT257R cloning vector. Following the confirmation of the cloned Luciferase transformed into E. coli. Recombinant clones were confirmed by specific PCR and restriction enzyme digestion analysis. The luxA and LuxB fragment was released subcloned in to the PcDNA3.1\hug and PcDNA3.1\neo expression vectors, respectively. recombinant plasmid was confirmed through restriction digestion using BamH1 and Kpn1 enzymes and subsequently, transformation procedure continued into NIH3T3 eukaryote cells by specific kit. Luminescence ability of recombinant clones was tested by NIH3T3 cells and dechanal (substrate) and Neomysin and hygromysin. The results showed that luminescence start after 2 hours and then increase after 6 hours. Inaddition, the protein identity was verified by western blot analysis, the protein bands 76 kD were detected. , which indicates protein expression of luxB, luxA.
    Description: Iranian Fisheries Science Research Institute
    Description: Published
    Keywords: Lusifrase ; Vibrio fischeri ; Transfer gene ; DNA ; Lusiferase gene ; Investihation ; Genetic ; Cyprins carpio ; Rutilus frisii kutum
    Repository Name: AquaDocs
    Type: Report , Refereed
    Format: 58pp.
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  • 2
    Publication Date: 2021-05-19
    Description: Over the past few years, the syndrome of streptococcosis has been associated with outbreaks in rainbow trout (Oncorhynchus mykiss, Walbaum) and caused significant economic losses in the aquaculture industry in Iran. The main purpose of this work was molecular identification of some causative agents of streptococcosis in rainbow trout. A total of 520 samples were collected from the head kidney of diseased fish (weight, 50_200g) in 72 farms of 8 provinces in Iran, during 2008 to 2009. Bacterial isolates representing morphology and biochemical profiles of Streptococcus spp. were further confirmed by polymerase chain reaction (PCR). DNA extraction was carried out from a single colony by using the extraction promega kit following the conditions described by the supplier. The PCR assay was developed based on the 16S rRNA and glucose kinase genes of Streptococcus spp. for the rapid and specific detection and identification of this pathogen from different sources. Approximately 40% of specimens were infected to Streptococcus spp. Consequently, five pathogenic species have been identified, including S. iniae in Fars province, S. faecium in Mazandaran province, S. agalactiae in Gilan and Mazandaran provinces, S. dysgalatiae in Lorestan, Kohgiluyeh and Boyerahmad, Gilan and Kermanshah provinces and S. uberis, which was common in all provinces (except Mazandaran and Lorestan). The dominant species (based on important species index) were S. uberis, S. dysgalactiae and S. agalactiae, respectively.
    Description: Published
    Keywords: Oncorhynchus mykiss ; Streptococcosis ; Rainbow trout ; Syndrome ; PCR ; DNA ; Distribution ; Molecular identification
    Repository Name: AquaDocs
    Type: Journal Contribution , Refereed
    Format: pp.109-122
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  • 3
    Publication Date: 2021-07-05
    Description: Over the past few years, the syndrome of streptococcosis has been associated with outbreaks in rainbow trout (Oncorhynchus mykiss, Walbaum) and caused significant economic losses in the aquaculture industry in Iran. The main purpose of this work was molecular identification of some causative agents of streptococcosis in rainbow trout. A total of 520 samples were collected from the head kidney of diseased fish (weight, 50_200g) in 72 farms of 8 provinces in Iran, during 2008 to 2009. Bacterial isolates representing morphology and biochemical profiles of Streptococcus spp. were further confirmed by polymerase chain reaction (PCR). DNA extraction was carried out from a single colony by using the extraction promega kit following the conditions described by the supplier. The PCR assay was developed based on the 16S rRNA and glucose kinase genes of Streptococcus spp. for the rapid and specific detection and identification of this pathogen from different sources. Approximately 40% of specimens were infected to Streptococcus spp. Consequently, five pathogenic species have been identified, including S. iniae in Fars province, S. faecium in Mazandaran province, S. agalactiae in Gilan and Mazandaran provinces, S. dysgalatiae in Lorestan, Kohgiluyeh and Boyerahmad, Gilan and Kermanshah provinces and S. uberis, which was common in all provinces (except Mazandaran and Lorestan). The dominant species (based on important species index) were S. uberis, S. dysgalactiae and S. agalactiae, respectively.
    Keywords: Aquaculture ; Biology ; Iran ; Streptococcosis ; Rainbow trout ; Syndrome ; PCR ; DNA ; Distribution ; S. uberis ; S. dysgalactiae ; S. agalactiae ; species ; fish
    Repository Name: AquaDocs
    Type: article , TRUE
    Format: application/pdf
    Format: application/pdf
    Format: 109-122
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  • 4
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    Iranian Fisheries Science Research Institute | Tehran, Iran
    In:  http://aquaticcommons.org/id/eprint/25639 | 18721 | 2018-10-08 05:54:55 | 25639 | Iranian Fisheries Science Research Institute
    Publication Date: 2021-07-16
    Description: In this study lusiferse gene extracted from Vibrio fischeri. V. fischeri strain was kindly provided by Iranian Research Organization for Science and Technology (IROST). Genomic DNA extraction was carried out by Phenol-chloroform. A DNA fragment encoding the luxA and LuxB was amplified by PCR using sense and antisense primers, specific restriction sites for BamH1 and Kpn1 were introduced into 5´ end of forward and reverse primer, respectively. The PCR product was purified from agarose gel and ligated into cleaved PT257R cloning vector. Following the confirmation of the cloned Luciferase transformed into E. coli. Recombinant clones were confirmed by specific PCR and restriction enzyme digestion analysis. The luxA and LuxB fragment was released subcloned in to the PcDNA3.1\hug and PcDNA3.1\neo expression vectors, respectively. recombinant plasmid was confirmed through restriction digestion using BamH1 and Kpn1 enzymes and subsequently, transformation procedure continued into NIH3T3 eukaryote cells by specific kit. Luminescence ability of recombinant clones was tested by NIH3T3 cells and dechanal (substrate) and Neomysin and hygromysin. The results showed that luminescence start after 2 hours and then increase after 6 hours. Inaddition, the protein identity was verified by western blot analysis, the protein bands 76 kD were detected which indicates protein expression of luxB, luxA.
    Keywords: Biology ; Iran ; Lusifrase ; Vibrio fischeri ; Transfer gene ; DNA ; Lusiferase gene ; Investihation ; Genetic ; Cyprins carpio ; Rutilus frisii kutum
    Repository Name: AquaDocs
    Type: monograph
    Format: application/pdf
    Format: application/pdf
    Format: 58
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  • 5
    Publication Date: 2021-07-16
    Description: In this study genetic characterization of Barbus brachycephalus caspius, Lucioperca lucioperca, Rutilus rutilus caspius, Rutilus frisi kutum and Salmo trutta caspius were examined by 611 samples from regions in East (Guilan province), Middle (Mazandaran province) and west (Golestan province) of southen part of the Caspian Sea. DNA was extracted from fin tissue by phenol -chlorophorm method and then PCR was performed using special primers. Statistical analysis of data was performed by Gene Alex, MEGA and Arlequin softwares. -Rutilus frisi kutum: The results showed that nine of ten primers were polymorphic loci. The mean of effective and observed alleles were 7.26±0.49 and 4.37±0.35 respectively. Also, the mean of expected and observed heterozygosity were 0.55±0.03 and 0.69±0.02 respectively Of the analysed loci, all of the samples (except Tajan and samples in LOC4 and Gilan samples in MFW2) possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium (P〈0.05). The genetic diversity was significantly different between samples of Golestan and Gilan, Golestan and sefidrood, Golestan and Tajan, Mazandaran and sefidrood and Gilan and Tajan (p〈0.05). -Rutilus rutilus caspius: Sevan variable microsatellite loci were used to investigate genetic diversity and population structure of R. rutilus caspius. The mean of effective and observed alleles were 5.75±0.30 and 4.76±0.25 respectively. Also, the mean of expected and observed heterozygosity were 0.58±0.18 and 0.73±0.01 respectively. All of the samples (except golestan samples in LOC3) possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium (P〈0.05) Of the analysed loci, the genetic divergence was significantly different between samples of Golestan and Gilan, Gilan and Mazandaran and Gilan with Gorgan bay (p〈0.05). -Salmo trutta caspius: Genetic characterization of S. trutta caspius was comparatively analyzed with mitochondrial DNA sequencing that 45 haplotypes was observed. The average of expected and observed heterozygosity were 0.61±0.35 and 0.33±0.12 respectively. The maximum of haplotype diversity (0.089±0.04) was in sardabrood river and the minimum was in Astara river (0.81±0.02). Also, the maximum of nucleotid diversity was 0.13±0.07 in Sardabrood and Chalos rivers and the minimum was 0.11±0.06 in Tonekabon river. In addition, the maximum and minimum of FST was 0.08 and 0.01 respectively. Of the analysed loci, the genetic divergence was significantly different between samples of Astara and Chalos, Astara and Tonekabon, Chalos and Karganrood and Tonekabon with Kaganrood (p〈0.05). - Barbus brachycephalus caspius: The size of amplified fragment was 800 bp in all of the samples. There were 24 variable loci and 12 haplotype that the maximum of haplotype was in Gilan area (8 haplotype). The average of expected and observed heterozygosity were 0.003±0.35 and 0.42±0.12 respectively. The results showed that the haplotype diversity was significantly different between samples of Sefidrood whit other samples (p〈0.05). In addition The maximum of nocleotid diversity was 0.005±0.003 in Sefidrood and minimum was 0.001± 0.001 in Tajan river. Of the analysed loci, the genetic divergence was significantly different between samples of Gilan and Tajan, Mazandaran and Sefidrood (p〈0.05). -Lucioperca lucioperca: The genetic diversity of L. lucioperca was analyzed by using microsatellite markers. Seven primer sequences available for were tested to amplify microsatellite loci that all of loci were polymorphic. The mean of effective and observed alleles were 6.14±0.45 and 3.88±0.34 respectively. Also, the mean of expected and observed heterozygosity were 0.662±0.03 and 0.70±0.02 respectively. The most of samples in PflaL6, PflaL7and PflaL8 loci possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium (P〈0.05). The maximum of FST was 0.30 between Gilan and Mazandaran samples that there were minimum gene flow (8.18). The genetic divergence was significantly different between samples of Gilan and Mazandaran and Golestan whit mazandaran (p〈0.05).
    Keywords: Biology ; Iran ; Caspian Sea ; Genetic variation ; Barbus brachycephalus caspius ; Lucioperca lucioperca ; Rutilus rutilus caspius ; Rutilus frisi kutum ; Salmo trutta caspius ; Identification ; Population ; Genetic ; DNA ; DNA bank ; Molecular ; Microsatellites
    Repository Name: AquaDocs
    Type: monograph
    Format: application/pdf
    Format: application/pdf
    Format: 82
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  • 6
    Publication Date: 2021-05-19
    Description: In this study genetic characterization of Barbus brachycephalus caspius, Lucioperca lucioperca , Rutilus rutilus caspius , Rutilus frisi kutum and Salmo trutta caspius were examined by 611 samples from regions in East (Guilan province), Middle (Mazandaran province) and west (Golestan province) of southen part of the Caspian Sea. DNA was extracted from fin tissue by phenol - chlorophorm method and then PCR was performed using special primers. Statistical analysis of data was performed by Gene Alex, MEGA and Arlequin softwares. - Rutilus frisi kutum: The results showed that nine of ten primers were polymorphic loci. The mean of effective and observed alleles were 7.26±0.49 and 4.37± 0.35 respectively. Also, the mean of expected and observed heterozygosity were 0.55±0.03 and 0.69±0.02 respectively. Of the analysed loci, all of the samples (except Tajan and samples in LOC4 and Gilan samples in MFW2) possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium(P〈0.05). The genetic diversity was significantly different between samples of Golestan and Gilan, Golestan and sefidrood, Golestan and Tajan, Mazandaran and sefidrood and Gilan and Tajan (p〈0.05). - Rutilus rutilus caspius: Sevan variable microsatellite loci were used to investigate genetic diversity and population structure of R. rutilus caspius. The mean of effective and observed alleles were 5.75±0.30 and 4.76± 0.25 respectively. Also, the mean of expected and observed heterozygosity were 0.58±0.18 and 0.73±0.01 respectively. All of the samples (except golestan samples in LOC3) possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium(P〈0.05). Of the analysed loci, the genetic divergence was significantly different between samples of Golestan and Gilan, Gilan and Mazandaran and Gilan with Gorgan bay (p〈0.05). - Salmo trutta caspius: Genetic characterization of S. trutta caspius was comparatively analyzed with mitochondrial DNA sequencing that 45 haplotypes was observed. The average of expected and observed heterozygosity were 0.61±0.35 and 0.33±0.12 respectively. The maximum of haplotype diversity (0.089±0.04) was in sardabrood river and the minimum was in Astara river (0.81±0.02). Also, the maximum of nucleotid diversity was 0.13±0.07 in Sardabrood and Chalos rivers and the minimum was 0.11±0.06 in Tonekabon river. In addition, the maximum and minimum of FST was 0.08 and 0.01 respectively. Of the analysed loci, the genetic divergence was significantly different between samples of Astara and Chalos, Astara and Tonekabon, Chalos and Karganrood and Tonekabon with Kaganrood (p〈0.05). - Barbus brachycephalus caspius: The size of amplified fragment was 800 bp in all of the samples. There were 24 variable loci and 12 haplotype that the maximum of haplotype was in Gilan area (8 haplotype). The average of expected and observed heterozygosity were 0.003±0.35 and 0.42±0.12 respectively. The results showed that the haplotype diversity was significantly different between samples of Sefidrood whit other samples (p〈0.05). In addition The maximum of nocleotid diversity was 0.005±0.003 in Sefidrood and minimum was 0.001± 0.001 in Tajan river. Of the analysed loci, the genetic divergence was significantly different between samples of Gilan and Tajan, Mazandaran and Sefidrood (p〈0.05). - Lucioperca lucioperca: The genetic diversity of L. lucioperca was analyzed by using microsatellite markers. Seven primer sequences available for were tested to amplify microsatellite loci that all of loci were polymorphic. The mean of effective and observed alleles were 6.14±0.45 and 3.88±0.34 respectively. Also, the mean of expected and observed heterozygosity were 0.662±0.03 and 0.70±0.02 respectively. The most of samples in PflaL6 , PflaL7and PflaL8 loci possible tests were found to deviate significantly from the Hardy–Weinberg equilibrium (P〈0.05). The maximum of FST was 0.30 between Gilan and Mazandaran samples that there were minimum gene flow (8.18). The genetic divergence was significantly different between samples of Gilan and Mazandaran and Golestan whit mazandaran (p〈0.05).
    Description: Iranian Fisheries Science Research Institute
    Description: Published
    Keywords: Genetic variation ; Barbus brachycephalus caspius ; Lucioperca lucioperca ; Rutilus rutilus caspius ; Rutilus frisi kutum ; Salmo trutta caspius ; Identification ; Population ; Genetic ; DNA ; DNA bank ; Molecular ; Microsatellites
    Repository Name: AquaDocs
    Type: Report , Refereed
    Format: 82pp.
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