• Title/Summary/Keyword: Fragment method

Search Result 555, Processing Time 0.027 seconds

Isolation and Identification of ura5 Gene in Entomopathogenic Fungus, Metarhizium anisopoliae (살충성곰팡이 Metarhizium anisopliae의 ura5 유전자의 분리동정)

  • Park, In-Cheol;Lee, Dong-Kyu;Kang, Sun-Cheol;Hwang, Cher-Won
    • Applied Biological Chemistry
    • /
    • v.40 no.1
    • /
    • pp.30-33
    • /
    • 1997
  • About 250 bp ura5 gene (Orotate phosphoribosyl transferase) fragment was cloned from genomic DNA of entomopathogenic fungus Metarhizium anisopliae by using PCR method. Entire nucleotide sequences of cloned DNA fragment were determined and analysed as compared with other fungus ura5 genes. The amino acid sequence deduced from the nucleotide sequence showed 85.5% homology to ura5 protein of Trichoderma reesei. Using this 250 bp PCR fragment we have isolated full ura5 gene of M. anisopliae by genomic Southern hybridization and the isolated 4.4 kb DNA fragments were mapped by restrictional enzyme.

  • PDF

Screening of BCL-2 associated X protein gene polymorphism associated with scrotal hernia in domesticated swine using polymerase chain reaction-restriction fragment length polymorphism

  • Manalaysay, Jessica G.;Antonio, Nathaniel D.;Apilado, Ralph Lorenz R.;Bambico, Joseph F.;Mingala, Claro N.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.30 no.2
    • /
    • pp.262-266
    • /
    • 2017
  • Objective: This study was conducted to screen scrotal hernia in domesticated swine from selected breeders in the Philippines. This defect is associated with a cytosine to thymine mutation in the BCL-2 associated X protein (BAX) gene of swine. Methods: Genetic screening was done by DNA extraction followed by amplification and digestion using polymerase chain reaction-restriction fragment length polymorphism, amplifying the 416 bp region of the BAX gene that was subjected to digestion using the Ear I enzyme. Sequencing was also conducted to validate the results. Results: Results revealed that out of 538 samples tested, 411 (76.4%) of the samples were found to be normal whereas the remaining were carriers of the mutation in which 80 (14.9%) were heterozygous mutants and 47 (8.7%) were homozygous mutants. Pietrain breed was found to have the highest incidence. Conclusion: Having a scrotal hernia eliminates the chances of using the boar as a breeder stock because the following generations arising from it would most likely exhibit herniation. It is therefore advised to establish a genetic screening method for Scrotal Hernia in the Philippines to eliminate the negative gene from the herd.

Comparison of Macroscopic Inspection and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) for the Detection of Anisakis simplex complex (고래회충 검출을 위한 육안검사법과 중합효소연쇄반응-제한효소절편길이다형성의 비교)

  • Kang, Ju-Hee;Lee, Min-Hwa;Lee, Kang-Bum;Choi, Chang-Sun
    • Journal of Food Hygiene and Safety
    • /
    • v.23 no.4
    • /
    • pp.314-318
    • /
    • 2008
  • This research aimed to compare the detection methods of Anisakis simplex in Sea fish by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and macroscopic inspection. We examined 18 Trichiurus lepturus, 11 Scomber japonicus, and 65 Todarodes pacificus collected from the retail markets in the areas of Uljin, Kyuonggi province and Seoul. As the result of examinations, we found that detection rate of Anisakis simplex by macroscopic observation was 89% in Trichiurus lepturus, 90.9% in Scomber japonicus, 32.3% in Todarodes pacificus. The detection rate of Anisakis simplex by PCR-RFLP was 77.7% in Trichiurus lepturus, 81.8% in Scomber japonicus, 26.1% in Todarodes pacificus. We could conclude that PCR-RFLP method of Anisakis simplex was more specific rather than macroscopic observation.

Numerical Analysis of Steel-strengthened Concrete Panels Exposed to Effects of Blast Wave and Fragment Impact Load Using Multi-solver Coupling (폭풍파 및 파편 충돌에 대한 강판보강 콘크리트 패널의 복합적 수치해석)

  • Yun, Sung-Hwan;Park, Taehyo
    • KSCE Journal of Civil and Environmental Engineering Research
    • /
    • v.31 no.1A
    • /
    • pp.25-33
    • /
    • 2011
  • The impact damage behavior of steel-strengthened concrete panels exposed to explosive loading is investigated. Since real explosion experiments require the vast costs to facilities as well as the blast and impact damage mechanisms are too complicated, numerical analysis has lately become a subject of special attention. However, for engineering problems involving blast wave and fragment impact, there is no single numerical method that is appropriate to the various problems. In order to evaluate the retrofit performance of a steel-strengthened concrete panel subject to blast wave and fragment impact loading, an explicit analysis program, AUTODYN is used in this work. The multi-solver coupling methods such as Euler-Lagrange and SPH-Lagrange coupling method in order to improve efficiency and accuracy of numerical analysis is implemented. The simplified and idealized two dimensional and axisymmetric models are used in order to obtain a reasonable computation running time. As a result of the analysis, concrete panels subject to either blast wave or fragment impact loading without the steel plate are shown the scabbing and perforation. The perforation can be prevented by concrete panels reinforced with steel plate. The numerical results show good agreement with the results of the experiments.

A chemical conjugate between HER2-targeting antibody fragment and Pseudomonas exotoxin A fragment demonstrates cytotoxic effects on HER2-expressing breast cancer cells

  • Lee, Sunju;Park, Sangsu;Nguyen, Minh Tan;Lee, Eunyoung;Kim, Julee;Baek, Sangki;Kim, Chong Jai;Jang, Yeon Jin;Choe, Han
    • BMB Reports
    • /
    • v.52 no.8
    • /
    • pp.496-501
    • /
    • 2019
  • Conventionally, immunotoxins have been produced as a single polypeptide from fused genes of an antibody fragment and a toxin. In this study, we adopted a unique approach of chemical conjugation of a toxin protein and an antibody fragment. The two genes were separately expressed in Escherichia coli and purified to high levels of purity. The two purified proteins were conjugated using a chemical linker. The advantage of this approach is its ability to overcome the problem of low recombinant immunotoxin production observed in some immunotoxins. Another advantage is that various combinations of immunotoxins can be prepared with fewer efforts, because the chemical conjugation of components is relatively simpler than the processes involved in cloning, expression, and purification of multiple immunotoxins. As a proof of concept, the scFv of trastuzumab and the PE24 fragment of Pseudomonas exotoxin A were separately produced using E. coli and then chemically crosslinked. The new immunotoxin was tested on four breast cancer cell lines variably expressing HER2. The chemically crosslinked immunotoxin exhibited cytotoxicity in proportion to the expression level of HER2. In conclusion, the present study revealed an alternative method of generating an immunotoxin that could effectively reduce the viability of HER2-expressing breast cancer cells. These results suggest the effectiveness of this method of immunotoxin crosslinking as a suitable alternative for producing immunotoxins.

Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • 2003.10a
    • /
    • pp.130-130
    • /
    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

  • PDF

Construction of Large Library of Protein Fragments Using Inter Alpha-carbon Distance and Binet-Cauchy Distance (내부 알파탄소간 거리와 비네-코시 거리를 사용한 대규모 단백질 조각 라이브러리 구성)

  • Chi, Sang-mun
    • Journal of the Korea Institute of Information and Communication Engineering
    • /
    • v.19 no.12
    • /
    • pp.3011-3016
    • /
    • 2015
  • Representing protein three-dimensional structure by concatenating a sequence of protein fragments gives an efficient application in analysis, modeling, search, and prediction of protein structures. This paper investigated the effective combination of distance measures, which can exploit large protein structure database, in order to construct a protein fragment library representing native protein structures accurately. Clustering method was used to construct a protein fragment library. Initial clustering stage used inter alpha-carbon distance having low time complexity, and cluster extension stage used the combination of inter alpha-carbon distance, Binet-Cauchy distance, and root mean square deviation. Protein fragment library was constructed by leveraging large protein structure database using the proposed combination of distance measures. This library gives low root mean square deviation in the experiments representing protein structures with protein fragments.

Isolation and Characterization of a Wound or UV Induced cDNA Fragment from Pleurotus sajor-caju (상처 및 자외선 자극에 반응하는 여름느타리 cDNA 단편의 분리 및 그 발현 특성)

  • Park, Soo-Chul;Jung, Uk-Jin;Jeong, Mi-Jeong;Kim, Bum-Gi;Yoo, Young-Bok;Ryu, Jin-Chang
    • The Korean Journal of Mycology
    • /
    • v.26 no.3 s.86
    • /
    • pp.314-320
    • /
    • 1998
  • A 0.4 kb cDNA fragment was isolated from mRNA of UV or mechanical wound damaged Pleurotus sajor-caju by the differential display method. Expression of the gene corresponding to this cDNA fragment was highly induced by mechanical wound damage or UV treatment. This gene showed only basal level expression in mycelia, stipe, and cap under normal growth conditions. Sequencing analysis showed that this cDNA fragment contains partial open reading frame. Homology search using genbank database revealed that although this gene do not have homology with already reported wound induced genes, it has a significant sequence homology in defined region with the cdc2-related protein kinase gene which is known to be involved in negative regulation of meiotic maturation in Xenopus oocytes.

  • PDF

Determinant Whether the Data Fragment in Unallocated Space is Compressed or Not and Decompressing of Compressed Data Fragment (비할당 영역 데이터 파편의 압축 여부 판단과 압축 해제)

  • Park, Bo-Ra;Lee, Sang-Jin
    • Journal of the Korea Institute of Information Security & Cryptology
    • /
    • v.18 no.4
    • /
    • pp.175-185
    • /
    • 2008
  • It is meaningful to investigate data in unallocated space because we can investigate the deleted data. However the data in unallocated space is formed to fragmented and it cannot be read by application in most cases. Especially in case of being compressed or encrypted, the data is more difficult to be read. If the fragmented data is encrypted and damaged, it is almost impossible to be read. If the fragmented data is compressed and damaged, it is very difficult to be read but we can read and interpret it sometimes. Therefore if the computer forensic investigator wants to investigate data in unallocated space, formal work of determining the data is encrypted of compressed and decompressing the damaged compressed data. In this paper, I suggest the method of analyzing data in unallocated space from a viewpoint of computer forensics.

A Study on Extracting the Document Text for Unallocated Areas of Data Fragments (비할당 영역 데이터 파편의 문서 텍스트 추출 방안에 관한 연구)

  • Yoo, Byeong-Yeong;Park, Jung-Heum;Bang, Je-Wan;Lee, Sang-Jin
    • Journal of the Korea Institute of Information Security & Cryptology
    • /
    • v.20 no.6
    • /
    • pp.43-51
    • /
    • 2010
  • It is meaningful to investigate data in unallocated space because we can investigate the deleted data. Consecutively complete file recovery using the File Carving is possible in unallocated area, but noncontiguous or incomplete data recovery is impossible. Typically, the analysis of the data fragments are needed because they should contain large amounts of information. Microsoft Word, Excel, PowerPoint and PDF document file's text are stored using compression or specific document format. If the part of aforementioned document file was stored in unallocated data fragment, text extraction is possible using specific document format. In this paper, we suggest the method of extracting a particular document file text in unallocated data fragment.