• 제목/요약/키워드: molecular sequence data

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SSR-Primer Generator: A Tool for Finding Simple Sequence Repeats and Designing SSR-Primers

  • Hong, Chang-Pyo;Choi, Su-Ryun;Lim, Yong-Pyo
    • Genomics & Informatics
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    • 제9권4호
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    • pp.189-193
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    • 2011
  • Simple sequence repeats (SSRs) are ubiquitous short tandem duplications found within eukaryotic genomes. Their length variability and abundance throughout the genome has led them to be widely used as molecular markers for crop-breeding programs, facilitating the use of marker-assisted selection as well as estimation of genetic population structure. Here, we report a software application, "SSR-Primer Generator " for SSR discovery, SSR-primer design, and homology-based search of in silico amplicons from a DNA sequence dataset. On submission of multiple FASTA-format DNA sequences, those analyses are batch processed in a Java runtime environment (JRE) platform, in a pipeline, and the resulting data are visualized in HTML tabular format. This application will be a useful tool for reducing the time and costs associated with the development and application of SSR markers.

Distinction between Cold-sensitive and -tolerant Jute by DNA Polymorphisms

  • Hossain, Mohammad Belayat;Awal, Aleya;Rahman, Mohammad Aminur;Haque, Samiul;Khan, Haseena
    • BMB Reports
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    • 제36권5호
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    • pp.427-432
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    • 2003
  • Jute is the principal coarse fiber for commercial production and use in Bangladesh. Therefore, the development of a high-yielding and environmental-stress tolerant jute variety would be beneficial for the agro economy of Bangladesh. Two molecular fingerprinting techniques, random-amplified polymorphic DNA (RAPD) and amplified-fragment length polymorphism (AFLP) were applied on six jute samples. Two of them were cold-sensitive varieties and the remaining four were cold-tolerant accessions. RAPD and AFLP fingerprints were employed to generate polymorphism between the cold-sensitive varieties and cold-tolerant accessions because of their simplicity, and also because there is no available sequence information on jute. RAPD data were obtained by using 30 arbitrary oligonucleotide primers. Five primers were found to give polymorphism between the varieties that were tested. AFLP fingerprints were generated using 25 combinations of selective-amplification primers. Eight primer combinations gave the best results with 93 polymorphic fragments, and they were able to discriminate the two cold-sensitive and four cold-tolerant jute populations. A cluster analysis, based on the RAPD and AFLP fingerprint data, showed the population-specific grouping of individuals. This information could be useful later in marker-aided selection between the cold-sensitive varieties and cold-tolerant jute accessions.

Expression of miR-210 during erythroid differentiation and induction of γ-globin gene expression

  • Bianchi, Nicoletta;Zuccato, Cristina;Lampronti, Ilaria;Borgatti, Monica;Gambari, Roberto
    • BMB Reports
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    • 제42권8호
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    • pp.493-499
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    • 2009
  • MicroRNAs (miRs) are a family of small noncoding RNAs that regulate gene expression by targeting mRNAs in a sequence specific manner, inducing translational repression or mRNA degradation. In this paper we have first analyzed by microarray the miR-profile in erythroid precursor cells from one normal and two thalassemic patients expressing different levels of fetal hemoglobin (one of them displaying HPFH phenotype). The microarray data were confirmed by RT-PCR analysis, and allowed us to identify miR-210 as an highly expressed miR in the erythroid precursor cells from the HPFH patient. When RT-PCR was performed on mithramycin-induced K562 cells and erythroid precursor cells, miR-210 was found to be induced in time-dependent and dose-dependent fashion, together with increased expression of the fetal $\gamma$-globin genes. Altogether, the data suggest that miR-210 might be involved in increased expression of $\gamma$-globin genes in differentiating erythroid cells.

Newly recorded species of the genus Synura (Synurophyceae) from Korea

  • Jo, Bok Yeon;Kim, Han Soon
    • Journal of Ecology and Environment
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    • 제41권1호
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    • pp.9-18
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    • 2017
  • Background: Species in the heterokont genus Synura are colonial and have silica scales whose ultrastructural characteristics are used for classification. We examined the ultrastructure of silica scales and molecular data (nuclear SSU rDNA and LSU rDNA, and plastid rbcL sequences) to better understand the taxonomy and phylogeny within the section Petersenianae of genus Synura. In addition, we report the first finding of newly recorded Synura species from Korea. Results: We identified all species by examination of scale ultrastructure using scanning and transmission electron microscopy (SEM and TEM). Three newly recorded species from Korea, Synura americana, Synura conopea, and Synura truttae were described based on morphological characters, such as cell size, scale shape, scale size, keel shape, number of struts, distance between struts, degree of interconnections between struts, size of base plate pores, keel pores, base plate hole, and posterior rim. The scales of the newly recorded species, which belong to the section Petersenianae, have a well-developed keel and a characteristic number of struts on the base plate. We performed molecular phylogenetic analyses based on sequence data from three genes in 32 strains (including three outgroup species). The results provided strong statistical support that the section Petersenianae was monophyletic, and that all taxa within this section had well-developed keels and a defined number of struts on the base plate. Conclusions: The phylogenetic tree based on sequence data of three genes was congruent with the data on scale ultrastructure. The resulting phylogenetic tree strongly supported the existence of the section Petersenianae. In addition, we propose newly recorded Synura species from Korea based on phylogenetic analyses and morphological characters: S. americana, S. conopea, and S. truttae.

Molecular characterization of a lectin, BPL-4, from the marine green alga Bryopsis plumosa (Chlorophyta)

  • Han, Jong-Won;Yoon, Kang-Sup;Jung, Min-Gui;Chah, Kyong-Hwa;Kim, Gwang-Hoon
    • ALGAE
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    • 제27권1호
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    • pp.55-62
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    • 2012
  • A novel lectin specific to N-acetyl-D-galactosamine as well as N-acetyl-D-glucosamine was isolated from Bryopsis plumosa and named as BPL-4. Sodium dodecyl sulfate polyacrylamide gel electrophorese (SDS-PAGE) and matrix-assisted laser desorption / ionization-time of flight (MALDI-TOF) mass spectrometry data showed that this lectin was a monomeric protein with molecular weight 12.9 kDa. The N-terminal amino acid sequences of the lectin were determined by Edman degradation and the full cDNA sequence encoding this lectin was obtained using the degenerate primers designed from the amino acid sequence. The size of the cDNA was 414 bp containing single open reading frame (ORF) encoding the lectin precursor. The homology analysis showed that this lectin might belong to H lectin group. BPL-4 showed high sequence similarity (60.6%) to BPL-3, which is a previously reported lectin from the same species. The comparative analysis on the lectin's primary structure showed two conserved domains including one possible active domain of H lectin group.

Improvement of Bacilysin Production in Bacillus subtilis by CRISPR/Cas9-Mediated Editing of the 5'-Untranslated Region of the bac Operon

  • Hadeel Waleed Abdulmalek;Ayten Yazgan-Karatas
    • Journal of Microbiology and Biotechnology
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    • 제33권3호
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    • pp.410-418
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    • 2023
  • Bacilysin is a dipeptide antibiotic composed of L-alanine and L-anticapsin produced by certain strains of Bacillus subtilis. Bacilysin is gaining increasing attention in industrial agriculture and pharmaceutical industries due to its potent antagonistic effects on various bacterial, fungal, and algal pathogens. However, its use in industrial applications is hindered by its low production in the native producer. The biosynthesis of bacilysin is mainly based on the bacABCDEF operon. Examination of the sequence surrounding the upstream of the bac operon did not reveal a clear, strong ribosome binding site (RBS). Therefore, in this study, we aimed to investigate the impact of RBS as a potential route to improve bacilysin production. For this, the 5' untranslated region (5'UTR) of the bac operon was edited using the CRISPR/Cas9 approach by introducing a strong ribosome binding sequence carrying the canonical Shine-Dalgarno sequence (TAAGGAGG) with an 8 nt spacing from the AUG start codon. Strong RBS substitution resulted in a 2.87-fold increase in bacilysin production without affecting growth. Strong RBS substitution also improved the mRNA stability of the bac operon. All these data revealed that extensive RBS engineering is a promising key option for enhancing bacilysin production in its native producers.

A Molecular Phylogenetic Study on Korean Alexandrium catenella and A. tamarense Isolates (Dinophyceae) Based on the Partial LSD rDNA Sequence Data

  • Kim, Keun-Yong;Kim, Chang-Hoon
    • Journal of the korean society of oceanography
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    • 제39권3호
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    • pp.163-171
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    • 2004
  • Sequences of the large subunit ribosomal (LSD) rDNA D1-D2 region of Alexandrium catenella(=A. sp. cf. catenella) and A. tamarense isolates, which were collected along the Korea coasts, were analyzed to understand their phylogenetic relationships and geographical distributions. All A. catenella and A. tamarense isolates belonged to the A. tamarense/catenella/fundyense complex and were grouped with the North American and temperate Asian ribotypes, respectively, regardless of the presence or absence of a ventral pore in the first apical plate. A consistent and peculiar characteristic that differentiated the Alexandrium isolates was amplification of a second PCR product with a lower molecular weight in addition to the predicted one; ten A. catenella isolates belonging to the temperate Asian ribotype yielded this additional PCR product. Sequence alignment revealed that the shorter PCR product resulted from an unusual large deletion of 87 bp in the LSD rDNA D1 domain. The North American and temperate Asian ribotypes were prevalent along the Korean coasts without geographical separation. Given the high genetic homogeneity among widely distributed Alexandrium populations, each ribotype appeared to be pandemic rather than to constitute a distinct regional population.

Refinements for the amplification and sequencing of red algal DNA barcode and RedToL phylogenetic markers: a summary of current primers, profiles and strategies

  • Saunders, Gary W.;Moore, Tanya E.
    • ALGAE
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    • 제28권1호
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    • pp.31-43
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    • 2013
  • This review provides a comprehensive summary of the PCR primers and profiles currently in use in our laboratory for red algal DNA barcoding and phylogenetic research. While work focuses on florideophyte taxa, many of the markers have been applied successfully to the Bangiales, as well as other lineages previously assigned to the Bangiophyceae sensu lato. All of the primers currently in use with their respective amplification profiles and strategies are provided, which can include full fragment, overlapping fragments and what might best be called "informed overlapping fragments", i.e., a fragment for a marker is amplified and sequenced for a taxon and those sequence data are then used to identify the best primers to amplify the remaining fragment(s) for that marker. We extend this strategy for the more variable markers with sequence from the external PCR primers used to "inform" the selection of internal sequencing primers. This summary will hopefully serve as a useful resource to systematists in the red algal community.

품질 정보와 퍼지 추론 기법을 이용한 DNA 염기 서열 배치 알고리즘 (A DNA Sequence Alignment Algorithm Using Quality Information and a Fuzzy Inference Method)

  • 김광백
    • 지능정보연구
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    • 제13권2호
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    • pp.55-68
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    • 2007
  • 분자 생물학(computational molecular biology) 분야에서 DNA 염기 서열 배치 알고리즘은 다양한 방법으로 개선되어 왔다. 본 논문에서는 기존의 DNA 염기의 품질 정보(quality information)를 이용한 DNA 염기 서열 배치 방법을 개선하기 위하여 퍼지 논리 시스템(fuzzy logic system)과 DNA 염기 서열 단편의 특징을 적용한 품질 정보와 퍼지 추론 기법을 이용한 DNA 염기 서열 배치 알고리즘을 제안한다. 기존의 알고리즘은 Needleman-Wunsch가 제안한 전역 배치 알고리즘에 각 DNA 염기의 품질 정보를 적용하여 DNA 염기 서열 배치 점수를 계산하였다. 그러나 전체 DNA 염기의 품질 정보를 이용하여 계산하기 때문에 DNA 염기 말단 부분의 품질이 낮은 경우에는 DNA 염기 서열 배치 점수를 계산하는 과정에서 오차가 발생한다. 본 논문에서는 기존의 품질 정보를 이용한 알고리즘을 개선하여 DNA 염기 서열의 말단 부위의 품질이 낮은 경우에도 정확히 서열을 배치할 수 있도록 한다. 또한 DNA 염기 서열 단편의 길이와 낮은 품질의 DNA 염기 빈도를 퍼지 논리 시스템에 적용하여 DNA 염기 서열 배치 점수를 계산하는데 적용되는 매핑 점수 인자(parameter)를 동적으로 조정한다. 제안된 알고리즘의 성능 평가를 위해 NCBI(National Center for Biotechnology Information)의 실체 유전체 데이터를 받아 성능을 분석한 결과, 제안된 알고리즘이 기존의 품질 정보만을 이용한 알고리즘 보다 DNA 염기 서열 배치에 있어서 효율적임을 확인하였다.

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