WordNet

in an unhelpful manner; "he stood by unhelpfully while the house burned down"
the quantity a skep can hold
an inability to be helpful

PrepTutorEJDIC

崇拝する,あがめる / 《the worshipful》《時にWorshipful》《英》《敬称として》尊敬すべき
(…の)一すくい分《+『of』+『名』》

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出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2016/01/29 20:14:43」(JST)

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Pfu DNA polymerase is an enzyme found in the hyperthermophilic archaeon Pyrococcus furiosus, where it functions to copy the organism's DNA during cell division. In the laboratory setting, Pfu is used to amplify DNA in the polymerase chain reaction (PCR), where the enzyme serves the central function of copying a new strand of DNA during each extension step.

Proofreading ability of Pfu polymerase

Pfu DNA polymerase has superior thermostability and proofreading properties compared to Taq DNA polymerase. Unlike Taq DNA polymerase, Pfu DNA polymerase possesses 3' to 5' exonuclease proofreading activity, meaning that as the DNA is assembled from the 5' end to 3' end, the exonuclease activity immediately removes nucleotides misincorporated at the 3' end of the growing DNA strand. Consequently, Pfu DNA polymerase-generated PCR fragments will have fewer errors than Taq-generated PCR inserts.

Commercially available Pfu typically results in an error rate of 1 in 1.3 million base pairs and can yield 2.6% mutated products when amplifying 1 kb fragments using PCR^{[citation needed]}. However, Pfu is slower and typically requires 1–2 minutes per cycle to amplify 1kb of DNA at 72 °C. Using Pfu DNA polymerase in PCR reactions also results in blunt-ended PCR products.

Pfu DNA polymerase is hence superior to Taq DNA polymerase for techniques that require high-fidelity DNA synthesis, but can also be used in conjunction with Taq polymerase to obtain the fidelity of Pfu with the speed of Taq polymerase activity.^{[citation needed]}.

History

Scientists associated with the biotech company Stratagene, based in La Jolla, California, discovered the superiority of Pfu over Taq in 1991. They published their work as Lundberg et al., "A High Fidelity Thermostable DNA Polymerase Isolated from Pyrococcus Furiosus," Strategies 4:34-35 (1991) and Gene in December of that year (Gene. 1991 Dec 12;108(1):1-6). U.S. Patent 5,489,523 was granted over exonuclease-deficient Pfu in February 1996 while U.S. and (1991)Patent 5,545,552 over Pfu itself was granted in August 1996.

External links

Stratagene's Pfu U.S. Patents
Patent 5,489,523
Patent 5,545,552

Molecular and Cellular Biology portal

UpToDate Contents

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1. Vaccination for the prevention chickenpox (primary varicella infection)
2. 遺伝学およびゲノミクスのためのツール：ポリメラーゼ連鎖反応 tools for genetics and genomics polymerase chain reaction
3. ムンプスの疫学、臨床症状、診断およびマネージメント epidemiology clinical manifestations diagnosis and management of mumps

English Journal

Generation and analysis of a large-scale expressed sequence tags from a full-length enriched cDNA library of Siberian tiger (Panthera tigris altaica).

Guo Y1, Liu C2, Lu T3, Liu D4, Bai C5, Li X6, Ma Y7, Guan W8.Author information 1Department of Bioscience, Department of Laboratory Medicine, Bengbu Medical College, Bengbu 233030, China. Electronic address: ily0720@126.com.2Department of Bioscience, Department of Laboratory Medicine, Bengbu Medical College, Bengbu 233030, China; Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: lcq7813@hotmail.com.3Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: taofenglu@yahoo.com.cn.4The Northeast Tiger Wooden Land of Heilongjiang, Harbin 150028, China. Electronic address: liudan_1964@sina.com.5Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; College of Wildlife Resource, Northeast Forestry University, Harbin 150028, China. Electronic address: baichunyu001@hotmail.com.6Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: nobelli@126.com.7Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: Yuehui.ma@263.net.8Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. Electronic address: wjguan86@hotmail.com.AbstractIn this study, a full-length enriched cDNA library was successfully constructed from Siberian tiger, the world's most endangered species. The titers of primary and amplified libraries were 1.28×10(6)pfu/mL and 1.59×10(10)pfu/mL respectively. The proportion of recombinants from unamplified library was 91.3% and the average length of exogenous inserts was 1.06kb. A total of 279 individual ESTs with sizes ranging from 316 to 1258bps were then analyzed. Furthermore, 204 unigenes were successfully annotated and involved in 49 functions of the GO classification, cell (175, 85.5%), cellular process (165, 80.9%), and binding (152, 74.5%) are the dominant terms. 198 unigenes were assigned to 156 KEGG pathways, and the pathways with the most representation are metabolic pathways (18, 9.1%). The proportion pattern of each COG subcategory was similar among Panthera tigris altaica, P. tigris tigris and Homo sapiens, and general function prediction only cluster (44, 15.8%) represents the largest group, followed by translation, ribosomal structure and biogenesis (33, 11.8%), replication, recombination and repair (24, 8.6%), and only 7.2% ESTs classified as novel genes. Moreover, the recombinant plasmid pET32a-TAT-COL6A2 was constructed, coded for the Trx-TAT-COL6A2 fusion protein with two 6× His-tags in N and C-terminal. After BCA assay, the concentration of soluble Trx-TAT-COL6A2 recombinant protein was 2.64±0.18mg/mL. This library will provide a useful platform for the functional genome and transcriptome research of for the P. tigris and other felid animals in the future.
Gene.Gene.2014 May 15;541(2):75-81. doi: 10.1016/j.gene.2014.03.023. Epub 2014 Mar 12.
In this study, a full-length enriched cDNA library was successfully constructed from Siberian tiger, the world's most endangered species. The titers of primary and amplified libraries were 1.28×10(6)pfu/mL and 1.59×10(10)pfu/mL respectively. The proportion of recombinants from unamplified library
PMID 24630959

Heat Shock Protein 70 Enhances Mucosal Immunity against Human Norovirus When Coexpressed from a Vesicular Stomatitis Virus Vector.

Ma Y1, Duan Y, Wei Y, Liang X, Niewiesk S, Oglesbee M, Li J.Author information 1Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.AbstractHuman norovirus (NoV) accounts for 95% of nonbacterial gastroenteritis worldwide. Currently, there is no vaccine available to combat human NoV as it is not cultivable and lacks a small-animal model. Recently, we demonstrated that recombinant vesicular stomatitis virus (rVSV) expressing human NoV capsid protein (rVSV-VP1) induced strong immunities in mice (Y. Ma and J. Li, J. Virol. 85:2942-2952, 2011). To further improve the safety and efficacy of the vaccine candidate, heat shock protein 70 (HSP70) was inserted into the rVSV-VP1 backbone vector. A second construct was generated in which the firefly luciferase (Luc) gene was inserted in place of HSP70 as a control for the double insertion. The resultant recombinant viruses (rVSV-HSP70-VP1 and rVSV-Luc-VP1) were significantly more attenuated in cell culture and viral spread in mice than rVSV-VP1. At the inoculation dose of 1.0 × 10(6) PFU, rVSV-HSP70-VP1 triggered significantly higher vaginal IgA than rVSV-VP1 and significantly higher fecal and vaginal IgA responses than rVSV-Luc-VP1, although serum IgG and T cell responses were similar. At the inoculation dose of 5.0 × 10(6) PFU, rVSV-HSP70-VP1 stimulated significantly higher T cell, fecal, and vaginal IgA responses than rVSV-VP1. Fecal and vaginal IgA responses were also significantly increased when combined vaccination of rVSV-VP1 and rVSV-HSP70 was used. Collectively, these data indicate that (i) insertion of an additional gene (HSP70 or Luc) into the rVSV-VP1 backbone further attenuates the VSV-based vaccine in vitro and in vivo, thus improving the safety of the vaccine candidate, and (ii) HSP70 enhances the human NoV-specific mucosal and T cell immunities triggered by a VSV-based human NoV vaccine.
Journal of virology.J Virol.2014 May;88(9):5122-37. doi: 10.1128/JVI.00019-14. Epub 2014 Feb 26.
Human norovirus (NoV) accounts for 95% of nonbacterial gastroenteritis worldwide. Currently, there is no vaccine available to combat human NoV as it is not cultivable and lacks a small-animal model. Recently, we demonstrated that recombinant vesicular stomatitis virus (rVSV) expressing human NoV cap
PMID 24574391

Development of a varicella-zoster virus neutralization assay using a glycoprotein K antibody enzyme-linked immunosorbent spot assay.

Chen L1, Liu J1, Wang W1, Ye J2, Wen L3, Zhao Q4, Zhu H5, Cheng T6, Xia N4.Author information 1State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China.2National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China.3Department of Obstetrics and Gynecology, the Affiliated Zhongshan Hospital of Xiamen University, Xiamen 361004, Fujian Province, China.4State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China; National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China.5Department of Microbiology and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, Newark, NJ 070101, USA.6State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, Xiamen 361102, Fujian Province, China; National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, Xiamen 361102, Fujian Province, China. Electronic address: tcheng@xmu.edu.cn.AbstractPlaque-reduction assays have been used to detect varicella-zoster virus (VZV)-neutralizing antibodies in sera for many decades. The current study characterized the mouse monoclonal antibody (MAb) 18A10, specific for VZV envelope glycoprotein K (gK), and applied this antibody to a new type of neutralization assay in the VZV field. The procedure is called the neutralization enzyme-linked immunosorbent spot (N-ELISPOT) assay and evolved from the VZV immunoperoxidase focus assay. Optimization of the assay involved defining the optimum combination of virus plaque-forming units (PFU) and antibody dilution, which were found to be 0-100PFU and 1:200, respectively. Furthermore, the N-ELISPOT assay produced results consistent with that obtained for the plaque-reduction neutralization assay. Considering that the plaque-reduction neutralization assay is time-consuming and labor-intensive, the VZV N-ELISPOT assay offers several advantages including reproducibility and applicability for high-throughput analysis of humoral immune responses to VZV.
Journal of virological methods.J Virol Methods.2014 May;200:10-4. doi: 10.1016/j.jviromet.2014.01.014. Epub 2014 Jan 31.
Plaque-reduction assays have been used to detect varicella-zoster virus (VZV)-neutralizing antibodies in sera for many decades. The current study characterized the mouse monoclonal antibody (MAb) 18A10, specific for VZV envelope glycoprotein K (gK), and applied this antibody to a new type of neutral
PMID 24486923

Japanese Journal

カーナビゲーションシステムの仮装走行環境によるテスト法

児島雄志,崔国東,DangVietHung ,森豪基,落水浩一郎
情報処理学会研究報告. ソフトウェア工学研究会報告 2011-SE-174(5), 1-8, 2011-10-25
現在,われわれはカーナビゲーションシステムの経路選択機能のためのテスト環境の構築に取り組んでいる.本稿では前述のテスト環境の構築において次の計算コストの節減のための 3 つの課題について報告する.1. テスト空間を適切に切り出すことでテストケースの数を節減する.2. カーナビゲーションシステムが選択した経路を評価するための比較対象経路群から重要ではない経路を排除する.3. 計算が単純で高い精度を持 …
NAID 110008668859

IaaS上でのContinuous Integrationによる運用支援フレームワークの提案

原田暢彦,山元隆弘,今井祐二,福井恵右
情報処理学会研究報告. [システムソフトウェアとオペレーティング・システム] 2011-OS-117(21), 1-9, 2011-04-06
ハードウェアおよびソフトウェアの組合せ結果として完成した業務システムは,運用開始後も機能追加や故障/不具合解消のために常に更新され続ける.クラウドの特性を活かして,更新される業務システムを自動ビルド&テストで全てのテストシナリオを実行することで,常に要求仕様を満足する機能提供を保証するシステム運用ツールを提案し,試作制作を通じて得た成果とシステム開発に課せられる課題を報告する.
NAID 110008583253