• Title/Summary/Keyword: Fragment

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Cloning of a Gene Involved in Biosynthesis of ${\beta}-1,3-glucan$ in Saccharomyces cerevisiae (베타-1,3-글루칸 생합성에 관여하는 Saccharomyces cerevisiae 유전자의 클로닝)

  • Jin, Eun-Hee;Lee, Dong-Won;Kim, Jin-Mi;Park, Hee-Moon
    • The Korean Journal of Mycology
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    • v.23 no.2 s.73
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    • pp.129-138
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    • 1995
  • DNA fragment being able to restore in vitro activity of ${\beta}-1,3-glucan$ synthase was cloned by transformation of the Saccharomyces cerevisiae LP353 mutant strain with genomic library constructed in the YCp50. For the selection of transformants which showed no detectable phenotype linked to recovery of the defect in ${\beta}-1,3-glucan$ synthase activity, the colony autoradiography was succesfully applied. The restriction map of the cloned DNA fragment, which is 8.5-kb in length, was constructed. Both the YEplac195 and the YCp50 carrying the 8.5-kb fragment increased ${\beta}-1,3-glucan$ synthase activity of LP353 by two fold. Neither the YEplac195 nor the YCp50 carrying the 8.5-kb DNA fragment, however, complemented the temperature-dependent osmotic sensitivity which is another distinctive phenotype of LP353. Subcloning experiments indicated that a functional region was located in 4.8-kb BglII-KpnI fragment. The 4.8-kb fragment was also able to increase the level of ${\beta}-1,3-glucan$ content in cell wall as well as the resistance of cells to cell wall lytic enzyme, ${\beta}-1,3-glucanase$. The growth rate of the LP353 with 4.8-kb fragment was almost same as that of wild type strain in liquid medium with 1.2 M sorbitol at nonpermissive temperature. Taken these results together, the 4.8-kb fragment seemed to contain the BGS2 gene for ${\beta}-1,3-glucan$ synthase activity in yeast S. cerevisiae.

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Genetic Differences and Variation in Two Largehead Hairtail (Trichiurus lepturus) Populations Determined by RAPD-PCR Analysis (RAPD-PCR 분석에 의해 결정된 갈치 (Trichiurus lepturus) 2 집단의 유전적 차이와 변이)

  • Park, Chang-Yi;Yoon, Jong-Man
    • Korean Journal of Ichthyology
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    • v.17 no.3
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    • pp.173-186
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    • 2005
  • Genomic DNA was isolated from two geographic populations of largehead hairtail (Trichiurus lepturus) in Korea and the Atlantic Ocean. The eight arbitrarily selected primers were found to generate common, polymorphic, and specific fragments. The complexity of the banding patterns varied dramatically between primers from the two locations. The size of the DNA fragments also varied widely, from 150 bp (base pairs) to 3,000 bp. Here, 947 fragments were identified in the largehead hairtail population from Korea, and 642 in the largehead hairtail population from the Atlantic Ocean: 148 specific fragments (15.6%) in the Korean population, and 61 (9.5%) in the Atlantic population. In the Korean population, 638 common fragments with an average of 79.8 per primer were observed.; 429 common fragments, with an average of 53.6 per primer, were identified in the Atlantic population. The number of polymorphic fragments in the largehead hairtail population from Korea and the Atlantic Ocean was 76 and 27, respectively. Based on the average bandsharing values of all samples, the similarity matrix ranged from 0.784 to 0.922 in the Korean population, and from 0.833 to 0.990 in the Atlantic population. The bandsharing value of individuals within the Atlantic population was much higher than in the Korean population. The dendrogram obtained by the eight primers indicated two genetic clusters: cluster 1 (KOREAN 01~KOREAN 11), and cluster 2 (ATLANTIC 12~ATLANTIC 22). Individual KOREAN no. 10 from Korea was genetically most closely related to KOREAN no. 11 in the Korean population (genetic distance = 0.038). Ultimately, individual KOREAN no. 01 of the Korean population was most distantly related to ATLANTIC no. 16 of the Atlantic population (genetic distance = 0.708).

Fragment Analysis for Detection of the FLT3-Internal Tandem Duplication: Comparison with Conventional PCR and Sanger Sequencing (FLT3-ITD 검출을 위한 절편분석법: 일반 중합효소연쇄반응 및 직접염기서열분석법과의 비교)

  • Lee, GunDong;Kim, Jeongeun;Lee, SangYoon;Jang, Woori;Park, Joonhong;Chae, Hyojin;Kim, Myungshin;Kim, Yonggoo
    • Laboratory Medicine Online
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    • v.7 no.1
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    • pp.13-19
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    • 2017
  • Background: We evaluated a sensitive and quantitative method utilizing fragment analysis of the fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD), simultaneously measuring mutant allele burden and length, and verified the analytical performance. Methods: The number and allelic burden of FLT3-ITD mutations was determined by fragment analysis. Serial mixtures of mutant and wild-type plasmid DNA were used to calculate the limit of detection of fragment analysis, conventional PCR, and Sanger sequencing. Specificity was evaluated using DNA samples derived from 50 normal donors. Results of fragment analysis were compared to those of conventional PCR, using 481 AML specimens. Results: Defined mixtures were consistently and accurately identified by fragment analysis at a 5% relative concentration of mutant to wild-type, and at 10% and 20% ratios by conventional PCR and direct sequencing, respectively. No false positivity was identified. Among 481 AML specimens, 40.1% (193/481) had FLT3-ITD mutations. The mutant allele burden (1.7-94.1%; median, 28.2%) and repeated length of the mutation (14-153 bp; median, 49 bp) were variable. The concordance rate between fragment analysis and conventional PCR was 97.7% (470/481). Fragment analysis was more sensitive than conventional PCR and detected 11 additional cases: seven had mutations below 10%, three cases represented conventional PCR failure, and one case showed false negativity because of short ITD length (14 bp). Conclusions: The new fragment analysis method proved to be sensitive and reliable for the detection and monitoring of FLT3-ITD in patients with AML. This could be used to simultaneously assess ITD mutant allele burden and length.

An Experimental Comparative Study of Radiography, Ultrasonography and CT Imaging in the IV Catheter Fragment (정맥내 카테터 조각의 엑스선, 초음파 및 CT 영상의 실험적 비교 연구)

  • Kweon, Dae Cheol
    • Journal of radiological science and technology
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    • v.39 no.2
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    • pp.185-191
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    • 2016
  • The objective of this study was to detect the fragments generated during IV (intravenous) catheter injection of contrast medium and drug administration in a clinical setting and removal was performed by experimentally producing a phantom, and to compare the radiography, ultrasonography, and multi-detector computed tomography (MDCT) imaging and radiation dose. A 1 cm fragment of an 18 gage Teflon$^{(R)}$ IV catheter with saline was inserted into the IV control line. Radiography, CT, and ultrasonography were performed and radiography and CT dose were calculated. CT and ultrasonography showed an IV catheter fragment clinically and radiography showed no visible difference in the ability to provide a useful image of an IV catheter fragment modality (p >.05). Radiography of effective dose ($0.2139mSv{\cdot}Gy^{-1}{\cdot}cm^{-2}$) form DAP DAP ($0.93{\mu}Gy{\cdot}m^2 $), and dose length product (DLP) ($201mGy{\cdot}cm$) to effective dose was calculated as 0.483 mSv. IV catheter fragment were detected of radiography, ultrasonography and CT. These results can be obtained by menas of an excellent IV catheter fragment of detection capability CT. However, CT is followed by radiation exposure. IV catheter fragment confirming the position and information recommend an ultrasonography.

Treatment of crown-root fracture with a modified crown fragment reattachment technique (변형된 치관부 파절편 재부착술식을 이용한 치관치근파절의 치료)

  • Song, Chang-Won;Song, Min-Ju;Shin, Su-Jung;Park, Jeong-Won
    • Restorative Dentistry and Endodontics
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    • v.35 no.5
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    • pp.395-400
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    • 2010
  • The development of adhesive dentistry has allowed that the crown fragment reattachment can be another option in the treatment of crown fracture. However, additional crown lengthening procedure or extrusion of the tooth may be necessary in the treatment of crown root fracture because subgingival fracture line in close proximity to the alveolar bone leads to challenges for restorative procedure and the violation of the biologic width. This case report presents a modified crown fragment reattachment technique of crown root fracture with pulp exposure, which was done without additional crown lengthening procedures. After the endodontic treatment, the patient was treated using a post insertion and the fragment reattachment technique, which made it possible to preserve the space for the biologic width and maintain a dry surgical field for adequate adhesion through the modification of the fractured coronal fragment. Since a coronal fracture was occurred and reattached afterward, it was observed that the coronal fragment was well maintained without the additional loss of periodontal attachment through 2-year follow up.

Long-Term Outcome of Reattached Tooth Fragment in Permanent Anterior Teeth of Children and Adolescents (소아 및 청소년의 영구치 치관 파절시 파절편 재부착술의 추적 관찰)

  • Kang, Hoyeon;Chae, Yongkwon;Lee, Koeun;Lee, Hyo-seol;Choi, Sungchul;Nam, Okhyung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.1
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    • pp.42-49
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    • 2021
  • This study aimed to evaluate the long-term outcomes of teeth treated with reattachment technique in children and adolescents. Twenty seven permanent anterior teeth from 21 patients treated with fragment reattachment were evaluated. Clinical photos and medical records were used to assess treatment outcomes. Effect of pulp treatment and the ratio of fragment on success rate were statistically analyzed. Detachment of fragment was observed in 17 teeth, and their duration of retention was 21.41 ± 23.39 months. Repeated trauma was found to be the most frequent causes of failure. Pulp treatment before reattachment did not affect the success rate (p > 0.05). The mean ratio of fragment was 0.482 ± 0.147, and the success rate was affected by the ratio of fragment (p = 0.018). The median retention time of the teeth was 72 months if the ratio was under 0.5, and 8 months for that of the others. A significant correlation was found between the ratio of fragment and retention time (p = 0.003). Reattachment can be a predictable treatment option for crown fracture in anterior teeth in children and adolescents when a fracture involves less than 50% of the clinical crown.

Investigation of the Copper (Cu) Binding Site on the Amyloid beta 1-16 (Aβ16) Monomer and Dimer Using Collision-induced Dissociation with Electrospray Ionization Tandem Mass Spectrometry

  • Ji Won Jang;Jin Yeong Lim;Seo Yeon Kim;Jin Se Kim;Ho-Tae Kim
    • Mass Spectrometry Letters
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    • v.14 no.4
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    • pp.153-159
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    • 2023
  • The copper ion, Cu(II), binding sites for amyloid fragment Aβ1-16 (=Aβ16 ) were investigated to explain the biological activity difference in the Aβ16 aggregation process. The [M+Cu+(z-2)H]z+ (z = 2, 3 and 4, M = Aβ16 monomer) and [D+Cu+(z-2)H]z+ (z = 3 and 5, D = Aβ16 dimer) structures were investigated using electrospray ionization (ESI) mass spectrometry (MS) and tandem mass spectrometry (MS/MS). Fragment ions of the [M+Cu+(z-2)H]z+ and [D+Cu+(z-2)H]z+ complexes were observed using collision-induced dissociation MS/MS. Three different fragmentation patterns (fragment "a", "b", and "y" ion series) were observed in the MS/MS spectrum of the (Aβ16 monomer or dimer-Cu) complex, with the "b" and "y" ion series regularly observed. The "a" ion series was not observed in the MS/MS spectrum of the [M+Cu+2H]4+ complex. In the non-covalent bond dissociation process, the [D+Cu+3H]5+ complex separated into three components ([M+Cu+H]3+, M3+, and M2+), and the [M+Cu]2+ subunit was not observed. The {M + fragment ion of [M+Cu+H]3+} fragmentation pattern was observed during the covalent bond dissociation of the [D+Cu +3H]5+ complex. The {M + [M+Cu+H]3+} complex geometry was assumed to be stable in the [D+Cu+3H]5+ complex. The {M + fragment ion of [M+Cu]2+} fragmentation pattern was also observed in the MS/MS spectrum of the [D+Cu+H]3+ complex. The {M + [y9+Cu]1+} fragment ion was the characteristic fragment ion. The [D+Cu+H]3+ and [D+Cu+3H]5+ complexes were likely to form a monomer-monomer-Cu (M-M-Cu) structure instead of a monomer-Cu-monomer (M-Cu-M) structure.

Calcaneal Apophyseal Avulsion Fractures with Achilles Tendon Rupture in a 10-Year-Old Patient: A Case Report (10세 남자 운동선수에서 발생한 아킬레스건 파열을 동반한 종골 골단의 견열 골절: 증례 보고)

  • Lee, Jun Young;Bak, Yi Gyu;Lim, Jae Hwan
    • Journal of Korean Foot and Ankle Society
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    • v.22 no.2
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    • pp.74-77
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    • 2018
  • Calcaneal apophysitis is a relatively common disease in young athletes. On the other hand, if not treated properly, it can lead to apophyseal avulsion fracture in rare cases. In the case of apophyseal avulsion fractures, it is often necessary to remove or preserve the bone fragment, which often requires a suture of the Achilles tendon. A 10-year-old badminton athlete visited the outpatients' clinic with pain in both heels from 10 months ago without any trauma history. After conservative therapy, the pain in the left heel was relived but the right heel pain persisted. After 10 months of conservative therapy, the patient visited the outpatients' clinic showing a calcaneal apophyseal avulsion fracture with a total rupture of the Achilles tendon. In the operation room, a bone fragment needed to be removed because of its poor viability and the fragment was too thin for fixation. After removing the bone fragment, the ruptured Achilles tendon was fixed with an anchor system.

A Study of the Anticoagulatory DNA from the Earthworm, Lumbricus rubellus, and its Regulatory DNA-Binding Protein

  • Kim, Gyoung-Mi;Yu, Kyoung-Hee;Woo, Jeong-Im;Bahk, Yun-Kyoung;Paik, Seung R.;Kim, Jung-Gyu;Chang, Chung-Soon
    • BMB Reports
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    • v.32 no.6
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    • pp.567-572
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    • 1999
  • We have previously shown that a DNA fragment is responsible for the anticoagulatory effect of an earthworm, Lumbricus rubellus. The anticoagluant increased the activated partial thromboplastin time (APTT) and also inhibited the thrombin activity observed with either N-${\alpha}$-p-tosyl-L-arginine methyl ester (TAME) or H-D-phenyl-alanyl-L-pipecoil-L-arginine-p-nitroanilide (S-2238). Since trypsin digestion of the anticoagulant further increased the APTT, the possible presence of a regulatory protein for the anticoagulatory DNA was investigated by digesting the anticoagulant with trypsin and isolating the DNA fragment with C4-reversed phase HPLC. The DNA fragment lacking a regulatory protein was eluted in the flow-through fraction, and analyzed with thrombin and activated factor X. Activated factor X activity was more strongly inhibited than thrombin activity. For DNA digestion, we treated the anticoagulant with DNase and purified the DNA-binding protein with a FPLC Resource-S cation exchange column. The regulatory protein, with an $M_r$ of 55.0 kDa, reduced the anticoagulatory effect of the DNA fragment.

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