• Title/Summary/Keyword: ITS1 sequencing

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Cloning and Sequencing Analysis of the Repressor Gene of Temperate Mycobacteriophage L1

  • Sau, Subrata;Chattoraj, Partho;Ganguly, Tridib;Lee, Chia Yen;Mandal, Nitai Chandra
    • BMB Reports
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    • v.37 no.2
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    • pp.254-259
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    • 2004
  • The wild-type and temperature-sensitive (ts) repressor genes were cloned from the temperate mycobacteriophage L1 and its mutant L1cIts391, respectively. A sequencing analysis revealed that the $131^{st}$ proline residue of the wild-type repressor was changed to leucine in the ts mutant repressor. The 100% identity that was discovered between the two DNA regions of phages L1 and L5, carrying the same sets of genes including their repressor genes, strengthened the speculation that L1 is a minor variant of phage L5 or vice versa. A comparative analysis of the repressor proteins of different mycobacteriophages suggests that the mycobacteriophage-specific repressor proteins constitute a new family of repressors, which were possibly evolved from a common ancestor. Alignment of the mycobacteriophage-specific repressor proteins showed at least 7 blocks (designated I-VII) that carried 3-8 identical amino acid residues. The amino acid residues of blocks V, VI, and some residues downstream to block VI are crucial for the function of the L1 (or L5) repressor. Blocks I and II possibly form the turn and helix 2 regions of the HTH motif of the repressor. Block IV in the L1 repressor is part of the most charged region encompassing amino acid residues 72-92, which flanks the putative N-terminal basic (residues 1-71) and C-terminal acidic (residues 93-183) domains of L1 repressor.

Molecular Identification of Asian Isolates of Medicinal Mushroom Hericium erinaceum by Phylogenetic Analysis of Nuclear ITS rDNA

  • Park, Hyuk-Gu;Ko, Han-Gyu;Kim, Seong-Hwan;Park, Won-Mok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.816-821
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    • 2004
  • A reliable molecular phylogenetic method to identify Hericium erinaceum, the most industrially valuable species in the Hericium genus, was established. Sequencing and phylogenetic analyses of the PCR-amplified ITS and 5.8S rDNA from Hericium fungi, including 6 species and 23 isolates, showed that variation in nucleotide sequences and size exists in both ITS1 and ITS2 regions, but not in the 5.8S region. These two ITS regions provided different levels of information on the relationship of H. erinaceum to other Hericium species. Based on the ITS1 sequence, both the parsimony and neighbor joining trees clearly distinguished Asian H. erinaceum isolates from other Hericium species and isolates. The intraspecific divergence of the ITS2 region was suitable to dissect the Asian H. erinaceum isolates into a few groups.

Isolation of Bacillus amyloliquefaciens Strains with Antifungal Activities from Meju

  • Lee, Hwang-A;Kim, Jeong-Hwan
    • Preventive Nutrition and Food Science
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    • v.17 no.1
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    • pp.64-70
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    • 2012
  • Bacilli with fibrinolytic activities were isolated from traditionally-prepared Meju and some of these strains showed strong antifungal activities. One isolate, MJ1-4, showed the strongest antifungal activity. MJ1-4 and other isolates were identified as B. amyloliquefaciens strains by recA gene sequencing and RAPD-PCR results. B. amyloliqufaciens MJ1-4 efficiently inhibited an Aspergillus spp.-producing aflatoxin B1 ($AFB_1$) and a Penicillium spp.-producing ochratoxin (OTA) in addition to other fungi. Antifungal activity of B. amyloliquefaciens MJ1-4 culture reached its maximum (40 AU/mg protein) in LB or TSB medium around 48 hr at $37^{\circ}C$. Antifungal activity of the concentrated culture supernatant was not decreased significantly by protease treatments, implying that the antifungal substance might not be a simple peptide or protein. Considering its antifungal and fibrinolytic activities together, B. amyloliquefaciens MJ1-4 can serve as a starter for fermented soyfoods such as Cheonggukjang and Doenjang.

Phylogenetic Analysis by RFLP and Sequencing of Mitochondrial DNA in a Korean Population

  • Lee, Jin-Young;Kim, Heui-Soo;Ha, Bae-Jin;Park, Yeong-Hong
    • Archives of Pharmacal Research
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    • v.29 no.1
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    • pp.88-95
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    • 2006
  • Analysis of molecular nature of mitochondrial DNA (mtDNA) could be powerful marker for anthropological studies of modern populations. While population genetic studies on mtDNA have been reported for several ethnic groups, no such study has been documented for the Korean population. We surveyed mtDNA polymorphisms in the HVS I of noncoding D-loop region and its upstream region from 430 unrelated healthy Korean population by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing analysis. PCR product with 2,790 bp spanning the specific mtDNA region (mt13715-16504) was subjected to RFLP analysis using 6 restriction enzyme (Hinf I, Hae III, Alu I, Dde I, Mbo I, Rsa I). On the PAUP analysis of PCR-RFLP results, 38 mtDNA haplotypes (Hap 1-38) were detected in the Korean populations, which were classified into 11 haplogroups (Grp 1-11) of related haplotypes encompassing all 38 haplotypes. In comparison of sequencing data with Anderson's reference sequence, the transition type was more prevalent than the transversion type. Insertions or deletions were not found. In addition, three of the polymorphic sites (A16240C, A16351G, G16384A) in HVS-I region are determined newly. The polymorphic sites were distributed randomly in the region, though the frequency at each site was variable. Thus, this research might be required for the genealogical study of Orientals.

A Study on Robustness Improvement of the Semiconductor Transmitter and Receiver Module By the Bias Sequencing and Tuning the Switching Time (바이어스 시퀀스와 스위칭 타임 튜닝을 통한 반도체 송수신 모듈의 강건성 향상에 대한 연구)

  • Yoo, Woo-Sung;Keum, Jong-Ju;Kim, Do-Yeol;Han, Sung
    • Journal of IKEEE
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    • v.20 no.3
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    • pp.251-259
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    • 2016
  • This paper describes that how to enhance the robustness of semiconductor TRM(Transmitter and Receiver Module) through the bias sequencing and tuning the switching time. Previous circuit designs focused on improving the MDS(Minimum Detection Signal) performance. Because TRM has critical problem which transmission output signal leak into receiver by it's compact design. Under this condition, TRM was frequently broken down within the MTBF(Mean Time Between Failure). This study proposes the bias sequencing and tuning the switching time to improve above problem. At first, we collected major failure symptom and infer it's cause. Second, we demonstrated it's effect by derive the improvement method and apply it to our system. And finally we can convinced that the proposed method clear the frequent failure problem with its lack of isolation.

Molecular Identification of Pooideae, Poaceae in Korea (국내 농경지에 발생하는 포아풀아과 잡초의 분자생물학적 동정)

  • Lee, Jeongran;Kim, Chang-Seok;Lee, In-Yong
    • Weed & Turfgrass Science
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    • v.4 no.1
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    • pp.18-25
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    • 2015
  • A universal DNA barcoding for agricultural noxious weeds is a powerful technique for species identification without morphological knowledge, by using short sections of DNA from a specific region of the genome. Two standard barcode markers, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used to examine the effectiveness of the markers for Pooideae barcoding using 163 individuals of 29 taxa across 16 genera of Korean Pooideae. The rbcL and ITS revealed a good level of amplification and sequencing success while matK did not. Barcode gaps were 78.6% for rbcL, 96.2% for matK, and 91.7% for ITS, respectively. Resolving powers were 89.3% for rbcL, 92.3% for matK, and 79.1% for ITS. The matK obtained the best both barcode gap and resolving power. However, it should be considered not to employ matK for Pooideae barcode because of low rate of PCR amplification and sequencing success. As a single DNA marker, rbcL and ITS were reasonable for Pooideae barcode. Barcode gap and resolving power were increased when ITS was incorporated into the rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use.

A Reliable "Direct from Field" PCR Method for Identification of Mycorrhizal Fungi from Associated Roots

  • Kuhnann, Christoph;Kim, Seak-Jin;Lee, Sang-Sun;Harms, Carsten
    • Mycobiology
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    • v.31 no.4
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    • pp.196-199
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    • 2003
  • A very reliable and specific method for the identification of fungi in ectotrophic mycorrhizal symbiosis was developed using a specific PCR assay based on the amplification of the ITS1 region. To obtain specific data, an ITS-diagnostic assay was carried out that reveals genera and species specific sequences. Here, an application of one method is presented, which covers the identification of pure mycelia, basidiocarps as well as mixed samples such as ectomycorrhizal roots that were mingled with remains of the host plant. For this purpose a protocol was established that allowed the extraction of DNA from single mycorrhizal roots. In order to perform a specific ITS analysis we generated a new ITS-primer(ITS8) by a multiple alignment of five different genera and species of mycorrhizal fungi. The utilization of ITS1 and ITS8 resulted in specific PCR amplicons, which were characterized by sequencing without purification steps, even when the template DNA was associated with roots.

Comparison of ITS(Internal Transcribed Spacer) and 5.8S rDNA Sequences among varieties and Cultivars in Panax ginseng

  • Yang, Deok-Chun;Yang, Key-Jin;Yoon, Eui-Soo
    • Journal of Photoscience
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    • v.8 no.2
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    • pp.55-60
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    • 2001
  • Ginseng (Panax genus) is one of the most medicinally important genera and consists of highly regarded medicines. Among the species of Panax, the ginseng species is widely known to have most medicinal quality. P. ginseng has 3 varieties, Jakyung, Chunggyung and Hwangsook, discovered in nature with different colors of stem and fruit, Jakyung has two cultivars, Yunpoong and Chunpoong. Rigorous phylogenetic analysis of these varieties and cultivars has been conducted with sequencing of rDNA region. The sequences of ITS1, ITS2 of every varieties and cultivars within P. ginseng were identical. The sequence of 5.8S rDNAs of Hwangsook variety were different from the sequences of 5.8S rDNAs of others by only one base pair at nucleotide position 14. In phylogenetic analysis and predicted RNA secondary structure study, it is assumed that evolution has proceeded from Hwangsook to other varieties. recently.

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Sequence Analysis of the Internal Transcribed Spacer of Ribosomal DNA in the Genus Rhizopus

  • Park, You-Jung;Min, Byung-Re
    • Mycobiology
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    • v.33 no.2
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    • pp.109-112
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    • 2005
  • The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with $154{\sim}155\;bp$. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. micros pores, R. homothallicus, and R. stolonifer groups.

Profiling Bartonella infection and its associated risk factors in shelter cats in Malaysia

  • Nurul Najwa Ainaa Alias;Sharina Omar;Nur Indah Ahmad;Malaika Watanabe;Sun Tee Tay;Nor Azlina Aziz;Farina Mustaffa-Kamal
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.38.1-38.12
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    • 2023
  • Background: Poor disease management and irregular vector control could predispose sheltered animals to disease such as feline Bartonella infection, a vector-borne zoonotic disease primarily caused by Bartonella henselae. Objectives: This study investigated the status of Bartonella infection in cats from eight (n = 8) shelters by molecular and serological approaches, profiling the CD4:CD8 ratio and the risk factors associated with Bartonella infection in shelter cats. Methods: Bartonella deoxyribonucleic acid (DNA) was detected through polymerase chain reaction (PCR) targeting 16S-23S rRNA internal transcribed spacer gene, followed by DNA sequencing. Bartonella IgM and IgG antibody titre, CD4 and CD8 profiles were detected using indirect immunofluorescence assay and flow cytometric analysis, respectively. Results: B. henselae was detected through PCR and sequencing in 1.0% (1/101) oral swab and 2.0% (1/50) cat fleas, while another 3/50 cat fleas carried B. clarridgeiae. Only 18/101 cats were seronegative against B. henselae, whereas 30.7% (31/101) cats were positive for both IgM and IgG, 8% (18/101) cats had IgM, and 33.7% (34/101) cats had IgG antibody only. None of the eight shelters sampled had Bartonella antibody-free cats. Although abnormal CD4:CD8 ratio was observed in 48/83 seropositive cats, flea infestation was the only significant risk factor observed in this study. Conclusions: The present study provides the first comparison on the Bartonella spp. antigen, antibody status and CD4:CD8 ratio among shelter cats. The high B. henselae seropositivity among shelter cats presumably due to significant flea infestation triggers an alarm of whether the infection could go undetectable and its potential transmission to humans.