• Title/Summary/Keyword: DNA Sequencing

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Sequence Verification of Synthetic Oligonucleotides by Exonuclease Digestion and Matrix Assisted Laser Desorption Ionization Mass Spectrometry

  • Kim, Jin-Sung;Jang, Jung-Suk;Choi, Jong-Soon;Chang, Yoon-Seok
    • BMB Reports
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    • v.29 no.2
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    • pp.122-126
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    • 1996
  • A series of oligonucleotides were synthesized by automatic DNA synthesizer. The purity of crude products was checked and their molecular weights determined by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) with an accuracy of better than 0.05% deviation even without using an internal standard. This mass determining technology in combination with partial digestion of oligonucleotides by 5'- and 3'-exonuclease provides a straightforward and simple method to obtain sequence information of oligonucleotides. The extension of this technology to the sequencing of modified oligonucleotides and genomic DNA and RNA might become possible.

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Molecular cloning and nucleotide sequencing of bovine viral diarrhea virus gp53 antigenic region (소 바이러스성 설사병 바이러스 gp53 항원부위 유전자의 재조합 및 염기서열 연구)

  • Yeo, Sang-geon;Cho, HJ;Masri, SA
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.287-295
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    • 1995
  • Molecular cloning and nucleotide sequencing were undertaken for the RNA genome of gp53 antigenic region in cytopathic Singer strain of bovine viral diarrhea virus. The cloned cDNA was 939 nucleotides in length having a base composition of 31.0% A, 19.6% C, 25.5% G and 24.0% T. The sequence was corresponded to approximately 77.8%(817 bases) of predicted gp53 region and 122 bases after 3'end of gp53 region in the Singer strain when compared with NADL strain of known sequence. A single open reading frame was found in the sequence of 2nd frame and was deduced as encoding 312 amino acids.

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Identification of Aspergillus nidulans from cooked eggs produced by permitted factory

  • Park, Hye-Won;Kim, Kook-Joo;Youk, Ji-Hea;Woo, Jong-Tae
    • Korean Journal of Veterinary Service
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    • v.30 no.2
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    • pp.283-286
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    • 2007
  • Fungus generally doesn't produce toxic or harmful substances so it has low chances to cause food poisoning. However it leads to change appearance, odor and characteristics of the contaminated foods and result in sanitary risk problems. Therefore the contamination of fungi should be prevented since they are not proper for human consumption. Green fungi with white outline raised from the air cell of cooked eggs which were collected by Gyeongi Livestock Veterinary Service in August, 2006. The results came out after the cultivation using Sabouraud's Dextrose Agar(SDA). The conidium appeared white and monospore, the shape of colony was round and oval. Conidiophore was brown and granulated and wrinkles and formed. It was confirmed as Aspergillus nidulans based on the dying using Lactophenol cotton blue, the observation of septum and vesicle from the grown spores, and rDNA sequencing.

Bridging Comparative Genomics and DNA Marker-aided Molecular Breeding

  • Choi, Hong-Kyu;Cook, Douglas R.
    • Korean Journal of Breeding Science
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    • v.43 no.2
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    • pp.103-114
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    • 2011
  • In recent years, genomic resources and information have accumulated at an ever increasing pace, in many plant species, through whole genome sequencing, large scale analysis of transcriptomes, DNA markers and functional studies of individual genes. Well-characterized species within key plant taxa, co-called "model systems", have played a pivotal role in nucleating the accumulation of genomic information and databases, thereby providing the basis for comparative genomic studies. In addition, recent advances to "Next Generation" sequencing technologies have propelled a new wave of genomics, enabling rapid, low cost analysis of numerous genomes, and the accumulation of genetic diversity data for large numbers of accessions within individual species. The resulting wealth of genomic information provides an opportunity to discern evolutionary processes that have impacted genome structure and the function of genes, using the tools of comparative analysis. Comparative genomics provides a platform to translate information from model species to crops, and to relate knowledge of genome function among crop species. Ultimately, the resulting knowledge will accelerate the development of more efficient breeding strategies through the identification of trait-associated orthologous genes and next generation functional gene-based markers.

Complete Genome Sequence of Levilactobacillus brevis KL251 Isolate from Kimchi

  • Kiyeop Kim;Da Jeong Shin;Junghee Lee;Sejong Oh
    • Journal of Dairy Science and Biotechnology
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    • v.42 no.1
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    • pp.18-22
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    • 2024
  • In this study, we performed whole-genome sequencing of Levilactobacillus brevis KL251 (KL251) isolated from kimchi. The KL251 genome, characterized by a circular chromosome spanning 2,345,062 base pairs with a GC content of 45.78%, was analyzed. KL251 contains 2,275 coding DNA sequences (CDSs), 56 transfer RNAs (tRNAs), and 4 ribosomal RNAs (rRNAs). Genes associated with gamma-aminobutyric acid (GABA) production and CRISPR-Cas systems were identified and could potentially be used for GABA synthesis and defense against foreign DNA. Additionally, the presence of functional genes involved in isoprenoid biosynthesis, glutathione generation, and redox sensing showed that cellular metabolism and stress responses were important characteristics of this genome. These genomic findings suggest that the KL251 strain could potentially have several applications, including food fermentation, probiotics, dairy product starters, and the development of health-enhancing products.

자리공 항바이러스 단백질 II 유전자의 형질전환에 의한 연초의 바이러스 저항성 품종 개발 (I)

  • 강신웅;이영기;이기원;박성원;이청호
    • Journal of the Korean Society of Tobacco Science
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    • v.21 no.1
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    • pp.57-63
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    • 1999
  • Pokeweed antiviral protein II (PAP-II) encoding cDNA was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from Phytolacca american a leaf. The PAP-II cDNA fragment of 974bp was subcloned to pBluescript II SK- SmaI site and the inserted PAP-II cDNA fragment was sequenced by dideoxy sequencing method. The number of nucleotides of PAP-II cDNA coding region containing start and stop codon was 933bp. To develop a virus-resistant tobacco plant, PAP-II cDNA fragment was inserted to pKGT101B and the insertion of PAP-II cDNA fragment was confirmed by restriction enzyme analysis and colony PCR.

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Characterization of the Nucleotide Sequence of a Polyubiquitin Gene (PUBC1) from Arabian Camel, Camelus dromedarius

  • Al-Khedhairy, Abdulaziz Ali A.
    • BMB Reports
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    • v.37 no.2
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    • pp.144-147
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    • 2004
  • Molecular amplification and sequencing of genomic DNA that encodes camel polyubiquitin (PUBC1) was performed by a polymerase chain reaction (PCR) using various sets of primers. The amplification generated a number of DNA fragments, which were sequenced and compared with the polyubiquitin coding sequences of various species. One DNA fragment that conformed to 325 bp was found to be 95 and 88% homologous to the sequences of human polyubiquitin B and C, respectively. The DNA translated into 108 amino acids that corresponded to two fused units of ubiquitin with no intervening sequence, which indicates that it is a polyubiquitin and contains at least two units of ubiquitin. Although, variations were found in the nucleotide sequence when compared to those of other species, the amino acid sequence was 100% homologous to the polyubiquitin sequences of humans, mice, and rats. This is the first report of the polyubiquitin DNA coding sequence and its corresponding amino acid sequence from camels, amplified using direct genomic DNA preparations.

Sequence analysis and cDNA probe hybridization of the nucleocapsid(N) protein gene of transmissible gastroenteritis virus(TGEV) and porcine epidemic diarrhea virus(PEDV) (Transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)의 nucleocapsid(N) 단백질 유전자에 대한 염기서열 분석과 cDNA probe hybridization)

  • Park, Ji-yong;Kim, Chul-joong;Shin, Kwang-soon;Kim, Won-yong;Kang, Shien-young;Park, Yong-ho;Han, Hae-jung;Park, Yong-ha
    • Korean Journal of Veterinary Research
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    • v.35 no.3
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    • pp.515-530
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    • 1995
  • Coronaviridae에 속하는 transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)를 specific하게 detection할 수 있는 방법을 개발하고자 본 연구를 수행하였다. 두 바이러스 모두 RNA 바이러스이기 때문에 reverse transcription-polymerase chain reaction(RT-PCR)으로 nucleocapsid(N) protein gene의 cDNA를 증폭시켰다. SmaI으로 처리한 pTZ19R에 ligation시킨 후 염기서열을 밝히고자 sequencing하였다. 각각의 prototype virus와 비교하여 상동성을 밝혔다. 두 바이러스에 대한 cDNA probe를 제작하여 Southern blot hybridization을 실시하였다. TGEV의 경우 백신주인 P45와 병독주인 Miller strain을 사용하였다. cDNA를 증폭시키기 위해 N1/N1R과 N2/N2R 두 가지 primer를 이용한 결과, N1/N1R primer의 경우 586bp 크기의 PCR product를 얻을 수 있었고, N2/N2R primers로 582bp의 cDNA를 증폭시킬 수 있었다. PEDV 실험을 위하여 PED 임상 증상을 나타내는 분변을 이용하여 RT-PCR을 실시하였다. P2/P2R primer로 753bp의 PCR product를 얻을 수 있었다. TGEV의 두 가지 strain의 N protein gene을 sequencing하여 prototype인 Purdue strain과 염기서열 상동성을 조사한 결과, 97%이상의 높은 homology를 나타내었다. PED-V 역시 N protein gene을 sequencing하여 CV777과 염기서열 상동성을 조사한 결과 97%이상의 homology로 PEDV임을 알 수 있었다. TGEV와 PEDV의 염기서열을 비교한 결과 29%의 낮은 homology를 관찰할 수 있었다. 두 가지 바이러스의 N protein gene에 대한 cDNA probe를 제작하여 Southern blot hybridization을 한 결과, 각 바이러스에 매우 특이적 반응을 나타내었다.

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Applications of Next-Generation Sequencing Technology based on Liquid Biopsy for Solid Tumor (고형암에서 액체 생검 기반의 차세대염기서열분석법 응용 기술)

  • Kim, Jin-Hee
    • Proceedings of the Korea Contents Association Conference
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    • 2019.05a
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    • pp.469-470
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    • 2019
  • 차세대염기서열분석법(NGS, Next Generation Sequencing) 기술은 하나의 유전체를 무수히 많은 조각으로 분해하여 각 조각을 동시에 읽어낸 뒤, 전산기술을 이용하여 조합함으로써 방대한 유전체 정보를 빠르게 해독하는 방법이다. 한편, 액체 생검(LB, liquid biopsy)이란 암세포가 깨지면서 생기는 미량의 DNA 조각을 말초혈액 속에서 찾아내 암을 진단하는 기술로 조직 생검(tissue biopsy)에 비해 비침습적이다. 본 논문은 NGS와 LB 기술을 접목했을 때 확진이 가능하고 예후 및 치료경과의 예측이 가능함을 제언하였다.

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