• 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.

• 한국발생생물학회지:발생과생식
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• 제16권4호
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• pp.379-384
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• 2012

Lhx8 (LIM homeobox 8) gene encodes a LIM homeodomain transcriptional regulator that is preferentially expressed in germ cells and critical for mammalian folliculogenesis. However, Lhx8 DNA binding sequences are not characterized yet. We aimed to identify and characterize a cis-acting sequence of germ-cell specific transcriptional factor, Lhx8. To identify Lhx8 DNA binding element, Cyclic Amplification of Sequence Target (CAST) Analysis was performed. Electrophoretic Mobility Shift Assay (EMSA) was processed for the binding specificity of Lhx8. Luciferase assay was for the transcriptional activity of Lhx8 through identified DNA binding site. We identified a putative cis-acting sequence, TGATTG as Lhx8 DNA binding element (LBE). In addition, Lhx8 binds to the LBE with high affinity and augments transcriptional activity of luciferase reporter driven by artificial promoter containing the Lhx8 binding element. These findings indicate that Lhx8 directly regulates the transcription of genes containing Lhx8 binding element in oocytes during early folliculogenesis.

• Journal of Microbiology
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• 제37권3호
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• pp.154-158
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• 1999

The arginine residue at position 243 (Arg 243) of the yeast transcription factor, Gcn4p, is invariably conserved among bZIP transcription factors. Using site-directed oligonucleotide saturation mutagenesis involving two-step polymerase chain reaction (PCR) amplification, random mutations were successfully introduced at the codon of 243 in the basic domain of Gcn4p. This mutant library was transformed ito Gcn4p defective yeast strain and selected for the transcriptionally active colonies. All colonies which were transcriptionally active had arginines in the codon 243. In this study, the strand preference by Taq polymerase during mutagenesis was also tested. Oligonucleotides were specially designed to test whether or not the polymerase was preferred using the strand as a template. A population of randomly mutated products were cloned into an appropriate vector and characterized by DNA sequencing analysis. Saturation mutagenesis which was performed efficiently by this method revealed a strong bias in terms of strand preference of Taq polymerase by an approximate ratio of 3 to 1 in this study.

• Journal of Microbiology and Biotechnology
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• 제8권1호
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• pp.34-41
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• 1998

A method was developed for the controlled expression of an antimicrobial peptide magainin in Escherichia coli. A series of concatemeric magainin genes was constructed with a gene amplification vector, and fused to the 3'end of malE gene encoding the affinity ligand, E. coli maltose-binding protein (MBP). The construct directed the synthesis of the fusion protein with the magainin polypeptide fused to the C-terminus of MBP. The fusion protein was expressed in a tightly regulatable expression system which was under the control of an invertible promoter. The MBP-fused magainin monomer was expressed efficiently. However, the expression level of the MBP-fused magainin in E. coli decreased with the increasing size of multimers possibly because of the transcription and translation inhibition by the multimeric peptides. After purification using an amylose affinity column, the fusion protein was digested by factor Xa at a specific cleavage site between the monomers. The recombinant magainin had an antimicrobial activity identical to that of synthetic magainin. This experiment shows that a biologically active, antimicrobial peptide magainin can be produced by fusing to MBP, along with a promoter inversion vector system.

• 한국지반공학회:학술대회논문집
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• 한국지반공학회 2008년도 춘계 학술발표회 초청강연 및 논문집
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• pp.1438-1445
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• 2008

The purpose of this study is to evaluate the fundamental frequency of the dam by the various methods using real microearthquake records which were measured on 'H' dam site and to compare each results. In this study, the fundamental frequency was evaluated by the frequency analysis of the microearthquake records which were measured on the dam crest, by the evaluation of acceleration amplification ratio between the foundation and the crest of dam, and by the evaluation of acceleration response spectrum ratio between the foundation and the crest of dam, respectively. Among these methods, it was found that the method by the evaluation of acceleration response spectrum ratio between the foundation and the crest of dam was the most effective method. But, if the simple engineering judgement can be considered, it was thought that the all three methods could reasonably evaluate the fundamental frequency of the dam.

• Journal of Crop Science and Biotechnology
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• 제10권4호
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• pp.237-242
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• 2007

Genome duplication(i.e. polyploidy) is a common phenomenon in the evolution of plants. The objective of this study was to achieve a comprehensive understanding of genome duplication for SNP discovery by Thymine/Adenine(TA) cloning for confirmation. Primer pairs were designed from 793 EST contigs expressed in the roots of a supernodulating soybean mutant and screened between 'Pureunkong' and 'Jinpumkong 2' by direct sequencing. Almost 27% of the primer sets were failed to obtain sequence data due to multiple bands on agarose gel or poor quality sequence data from a single band. TA cloning was able to identify duplicate genes and the paralogous sequences were coincident with the nonspecific peaks in direct sequencing. Our study confirmed that heterogeneous products by the co-amplification of a gene family member were the main cause of obtaining multiple bands or poor quality sequence data in direct sequencing. Counts of amplified bands on agarose gel and peaks of sequencing trace suggested that almost 27% of nonrepetitive soybean sequences were present in as many as four copies with an average of 2.33 duplications per segment. Copy numbers would be underestimated because of the presence of long intron between primer binding sites or mutation on priming site. Also, the copy numbers were not accurately estimated due to deletion or tandem duplication in the entire soybean genome.

• 전기학회논문지
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• 제65권5호
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• pp.857-861
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• 2016

In this study, we proposed a new method that can be measure ECG (Electrocardiography) and PPG (Photoplethysmography) in realtime on the site of the wrist for check the state of health in daily life. For convenience measurement of ECG the lead I method was used on the wrist, and omit the reference junction ECG I was measured in the right hand and the left hand of the potential difference. Then the measured electrocardiogram was amplified by the differential amplifier and the signals were passed HPF, LPF, and BPF filters. For removing the PPG's noise from the Motion artifact and temperature, we apply the reflective photoelectric volume pulse wave measurement method using green LED as a light source. The circuits was designed to be able to check the waveform using higher active amplification method at weak signals. For the validation of our device, the measured signals were compared with E2-KIT on same time. The results shows that the error does not exceed the maximum one, most of the data is confirmed to be issued Peak inspection of the same number.

• Bulletin of the Korean Chemical Society
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• 제25권6호
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• pp.833-837
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• 2004

DeniLite$^{TM}$ laccase immobilized platinum electrode was used for amperometric detection of hydroquinone (HQ) and homogentisic acid (HGA) by means of substrate recycling. In case of HQ, the obtained sensitivity is 280 nA/ ${\mu}$M with linear range of 0.2-35 ${\mu}$M ($r^2$ = 0.998) and detection limit (S/N = 3) of 50 nM. This high sensitivity can be attributed to chemical amplification due to the cycling of the substrate caused by enzymatic oxidation and following electrochemical regeneration. In case of HGA, the obtained sensitivity is 53 nA/ ${\mu}$M with linear range of 1-50 $[\mu}M\;(r^2$ = 0.999) and detection limit of 0.3 ${\mu}$M. The response times ($t_{90%}$) are about 2 seconds for the two substrates and the long-term stability is 60 days for HQ and around 40-50 days for HGA with retaining 80% of initial activities. The very fast response and the durable long-term stability are the principal advantages of this sensor. pH studies show that optimal pH of the sensor for HQ is 6.0 and that for HGA is 4.5-5.0. This shift of optimal pH towards acidic range for HGA can be attributed to the balance between enzyme activity and accessibility of the substrate to the active site of the enzyme.

• 보존과학연구
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• 통권23호
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• pp.59-75
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• 2002

In this study human bones and teeth, excavated from the Myeongam-risite in Asan, Chungcheongnam-do Province, have been analysed by nuclear DNA typing and mitochondrial DNA sequencing methods. Twenty-one samples of long bones and twenty-seven samples of teeth from twenty-one individuals were collected and analysed. Among these thirteenteeth were successfully subjected to nuclear DNA extraction, quantification, and PCR(Polymerase Chain Reaction) amplification. Silver STR III (D16S539, D7S820, D13S317) multiplex PCR method was used in this study for a short tandem repeat (STR) analysis. Mitochondrial DNAs of tooth samples were also amplified and sequenced by a DNA sequencer. These analyses show that a sample from Burial no. 29 and one from Burial no. 38(right) possessed the same maternal inheritance. This may suggest that the Myeongam-ri cemetery was used by a kin group for a relatively long period of time.

• Horticulture, Environment, and Biotechnology : HEB
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• 제59권6호
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• pp.857-864
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• 2018

Previous studies have not detected transcripts of the gene encoding anthocyanidin synthase (ANS) in white pomegranates (Punica granatum L.) and suggest that a large-sized insertion in the coding region of the ANS gene might be the causal mutation. To elucidate the identity of the putative insertion, 3887-bp 5' and 3392-bp 3' partial sequences of the insertion site were obtained by genome walking and a gene coding for an expansin-like protein was identified in these genome-walked sequences. An identical protein (GenBank accession OWM71963) isolated from pomegranate was identified from BLAST search. Based on information of OWM71963, a 5.8-Mb scaffold sequence with genes coding for the expansin-like protein and ANS were identified. The scaffold sequence assembled from a red pomegranate cultivar also contained all genome-walked sequences. Analysis of positions and orientations of these genes and genome-walked sequences revealed that the 27,786-bp region, including the 88-bp 5' partial sequences of the ANS gene, might be translocated into an approximately 22-kb upstream region in an inverted orientation. Borders of the translocated region were confirmed by PCR amplification and sequencing. Based on the translocation mutation, a simple PCR codominant marker was developed for efficient genotyping of the ANS gene. This molecular marker could serve as a useful tool for selecting desirable plants at young seedling stages in pomegranate breeding programs.