• IMMUNE NETWORK
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• 제2권2호
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• pp.79-85
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• 2002

Background: In contrast to evidences of Ig H chain receptor editing in transformed cell lines and transgenic mouse models, there has been no direct evidence that this phenomenon occurs in human developing B cells. Methods: $V_HDJ_H$ rearrangements were obtained from genomic DNA of individual $IgM^-$ B cells from liver and $IgM^+B$ cells from bone marrow of 18 wk of gestation human fetus by PCR amplification and direct sequencing. Results: We found three examples of H chain receptor editing from $IgM^+$ and $IgM^-human$ fetal B cells. Two types of $V_H$ replacements were identified. The first involved $V_H$ hybrid formation, in which part of a $V_H$ gene from the initial VDJ rearrangement is replaced by part of an upstream $V_H$ gene at the site of cryptic RSS. The second involved a gene conversion like replacement of CDR2, in which another $V_H$ gene donated a portion of its CDR2 sequence to the initial VDJ rearrangement. Conclusion: These data provide evidence of receptor editing at the H chain loci in developing human B cells, and also the first evidence of a gene conversion event in human Ig genes.

• Genomics & Informatics
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• 제18권1호
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• pp.6.1-6.9
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• 2020

Acute leukemia represents the most common pediatric malignancy comprising diverse subtypes with varying prognosis and treatment outcomes. New and targeted treatment options are warranted for this disease. Patient-derived xenograft (PDX) models are increasingly being used for preclinical testing of novel treatment modalities. A novel approach involving targeted error-corrected RNA sequencing using ArcherDX HemeV2 kit was employed to compare 25 primary pediatric acute leukemia samples and their corresponding PDX samples. A comparison of the primary samples and PDX samples revealed a high concordance between single nucleotide variants and gene fusions whereas other complex structural variants were not as consistent. The presence of gene fusions representing the major driver mutations at similar allelic frequencies in PDX samples compared to primary samples and over multiple passages confirms the utility of PDX models for preclinical drug testing. Characterization and tracking of these novel cryptic fusions and exonal variants in PDX models is critical in assessing response to potential new therapies.

• ALGAE
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• 제32권2호
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• pp.155-160
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• 2017

Red rot disease has caused a major decline in Pyropia (Nori) crop production in Korea, Japan, and China. To date, only Pythium porphyrae (Pythiales, Oomycetes) has been reported as the pathogen causing red rot disease in Pyropia yezoensis (Rhodophyta, Bangiales). Recently, Pythium chondricola was isolated from the infected blades of Py. yezoensis during molecular analyses using the mitochondrial cox1 region. In this study, we evaluated the pathogenicity of P. chondricola as an algal pathogen of Py. yezoensis. Moreover, a new cox2 marker was developed with high specificity for Pythium species. Subsequent to re-inoculation, P. chondricola successfully infected Py. yezoensis blades, with the infected regions containing symptoms of red rot disease. A novel cox2 marker successfully isolated the cox2 region of Pythium species from the infected blades of Py. yezoensis collected from Pyropia aquaculture farms. cox2 sequences showed 100% identity with that of P. chondricola (KJ595354) and 98% similarity with that of P. porphyrae (KJ595377). The results of the pathogenicity test and molecular analysis confirm that P. chondricola is a new algal pathogen causing red rot disease in Pyropia species. Moreover, it could also suggest the presence of cryptic biodiversity among Korean Pythium species.

• ALGAE
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• 제33권1호
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• pp.69-84
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• 2018

We examined the species diversity of Herposiphonia on Korean coasts, based on a combination of morphology and molecular analyses of the mitochondrial COI-5P DNA barcode marker and plastid rbcL gene. We report the presence of eight species including three novel species: H. donghaensis sp. nov., H. jejuinsula sp. nov., H. sparsa sp. nov., H. caespitosa, H. fissidentoides, H. insidiosa, H. parca, and H. subdisticha. Specimens were separated into eight clades in both the COI-5P and rbcL gene analyses, with 1.3-19.6 and 6.6-15% interspecific sequence divergence, respectively. These eight species are also distinguishable by several morphological characteristics such as: branching pattern (d/i pattern in H. donghaensis sp. nov. and H. sparsa sp. nov.; d/d/d/i pattern in others), shape of determinate branch (ligulate in H. fissidentoides; terete in others), number of vegetative trichoblasts (1-2 in H. insidiosa and H. sparsa sp. nov.; 3-4 in H. caespitosa; absent in others), and number of segments and pericentral cells in determinate branches. About three novel species revealed by our analyses, H. donghaensis sp. nov. is newly discovered, and H. jejuinsula sp. nov. and H. sparsa sp. nov. were previously reported in Korea as H. nuda and H. secunda, respectively. Our results show that DNA barcoding and rbcL analyses are useful for delimiting species boundaries and discovering cryptic species diversity in the genus Herposiphonia.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1986년도 추계학술대회
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• pp.529.2-529
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• 1986

pSp-Si (1.6kbp) was originally found in pediococcus halophilus to be a cryptic multicopy-plasmid. Hoping that the plasmid can also replicate in Bacillus subtilis, protoplast transformation of strain 207-25 (recE) was performed using pSP-Sl onto which was added the marker of tmrB8 (on 4.9 kbp EcoRI fragment ) or tmrB+ (on 0.9 kbp xbaI fragment) gene. Though the tmrB8 gene can expres tunicamycin-resistance at the single copy state, and the tmrB+ gene exerts the resistance only at the multicopy state, we could not confirm the replication of pSP-Sl (tmrB8) or pSP-Sl(tmrB+) in B. subtilis. During the experiment, however, we unexpectedly found that the circularized 0.9 kbp xgaI fragment (tmrB+) itself, which had no replication origin, could transform strain 207-25 to tunicamycin-resistant by protoplast transformation. Southern hybridization analyses with tmrB+ and other probes revealed the integration of the fragment at a single copy state into a position other than the homologous tmrB gene. This recE independent integration of another tmrB+ gene into the chromosome may contribute to the tunicamycinresistance in the transformants.

• Journal of Microbiology and Biotechnology
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• 제29권12호
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• pp.1931-1937
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• 2019

The heterologous expression of the Streptomyces natural product (NP) biosynthetic gene cluster (BGC) has become an attractive strategy for the activation, titer improvement, and refactoring of valuable and cryptic NP BGCs. Previously, a Streptomyces artificial chromosomal vector system, pSBAC, was applied successfully to the precise cloning of large-sized polyketide BGCs, including immunosuppressant tautomycetin and antibiotic pikromycin, which led to stable and comparable production in several heterologous hosts. To further validate the pSBAC system as a generally applicable heterologous expression system, the daptomycin BGC of S. roseosporus was cloned and expressed heterologously in a model Streptomyces cell factory. A 65-kb daptomycin BGC, which belongs to a non-ribosomal polypeptide synthetase (NRPS) family, was cloned precisely into the pSBAC which resulted in 28.9 mg/l of daptomycin and its derivatives in S. coelicolor M511(a daptomycin non-producing heterologous host). These results suggest that a pSBAC-driven heterologous expression strategy is an ideal approach for producing low and inconsistent Streptomyces NRPS-family NPs, such as daptomycin, which are produced low and inconsistent in native host.

• Asian Pacific Journal of Cancer Prevention
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• 제17권2호
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• pp.553-555
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• 2016

Background: Liver cytology is indicated and requested for evaluating hepatic masses in symptomatic or serendipitous cryptic discovered lesions. Objective: To determine the cytomorphological patterns of hepatic lesions identified among a group of Sudanese patients. Materials and Methods: This is an analytical descriptive hospital-based study included 165 patients undergoing ultrasound-guided fine needle is an aspiration cytology (FNAC)for hepatic lesions, at Al-Amal Military Hospital & Khartoum Teaching Hospital in Khartoum, Sudan. Clinical data were reviewed. Air dried Diff Quick stained smears were grouped into unsatisfactory samples, benign lesions, and malignant neoplasms. Results: Our population were consisted of 35 (21.2%) females and 130 (78.8%) males, with a male to female ratio 3.7:1 and an age ranged between 47 to 80, and a mean age $57{\pm}7$. Of 165 cases, 57 (34.5%) were benign, no atypia were noticed, 101 (61.2%) were malignant. Most investigated patients were found to have metastatic lesions. Conclusion: FNAC is a useful tool for investigating hepatic lesions.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.164-169
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• 1995

An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.

• ALGAE
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• 제30권1호
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• pp.15-33
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• 2015

When the generitype Centroceras clavulatum, a presumed cosmopolitan warm temperate to tropical red alga, was discovered to have a biogeographic distribution limited to the Pacific Ocean using molecular and morphological evidence, the taxonomy in the genus Centroceras was thrown into chaos worldwide. An analysis of what species was, or were, previously identified as C. clavulatum in Bermuda is the focus of the present molecular (COI-5P, rbcL) and morphological study. Two novel species are proposed, C. arcii sp. nov. and C. illaqueans sp. nov., and the distributions of three taxa recently segregated in the 'C. clavulatum complex' of the western Atlantic, C. gasparrinii, C. hyalacanthum, and C. micracanthum, have been expanded to include Bermuda. C. arcii is shown to be morphologically cryptic with C. micracanthum, and remains best distinguished by its COI-5P barcode sequence.

• Animal Systematics, Evolution and Diversity
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• 제27권2호
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• pp.159-163
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• 2011

DNA barcoding of Bryozoa or "moss animals" has hardly advanced and lacks reference sequences for correct species identification. To date only a small number of cytochrome c oxidase subunit I (COI) sequences from 82 bryozoan species have been deposited in the National Center for Biotechnology Information (NCBI) GenBank and Barcode of Life Data Systems (BOLD). We here report COI data from 53 individual samples of 29 bryozoan species collected from Korean seawater. To our knowledge this is the single largest gathering of COI barcode data of bryozoans to date. The average genetic divergence was estimated as 23.3% among species of the same genus, 25% among genera of the same family, and 1.7% at intraspecific level with a few rare exceptions having a large difference, indicating a possibility of presence of cryptic species. Our data show that COI is a very appropriate marker for species identification of bryozoans, but does not provide enough phylogenetic information at higher taxonomic ranks. Greater effort involving larger taxon sampling for the barcode analyses is needed for bryozoan taxonomy.