• Title/Summary/Keyword: molecular bands

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Differentially Expressed Genes in Hemocytes of Vibrio harveyi-challenged Shrimp Penaeus monodon

  • Somboonwiwat, Kunlaya;Supungul, Premruethai;Rimphanitchayakit, Vichien;Aoki, Takashi;Hirono, Ikuo;Tassanakajon, Anchalee
    • BMB Reports
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    • v.39 no.1
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    • pp.26-36
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    • 2006
  • Differential Display PCR technique (DD-PCR) was used for the analysis of altered gene expression in hemocytes of Vibrio harveyi-infected Penaeus monodon. Forty-four combinations of arbitrary and oligo(dT) primers were used to screen for differentially expressed genes. A total of 79 differentially expressed bands could be identified from 33 primer combinations. These included 48 bands (61%) whose expression level increased and 31 bands (39%) decreased after V. harveyi challenge. Subsequently, forty-eight differential display fragments were successfully reamplified and cloned. A total of 267 clones were randomly selected and sequenced. The sequence analysis showed that 85 (31%) out of 267 clones were matched with sequences in the GenBank database which represented 24 different genes with known functions. Among the known genes, glucose transporter 1, interferon-related developmental regulator 1, lysozyme, profilin, SERPINB3, were selected for further confirmation of their differentially expression patterns by real-time PCR. The results showed increasing in expression level of the selected genes in shrimp hemocytes after microbial challenge suggesting the involvement of such genes in bacterial response in shrimp. The anti-lipopolysaccharide factor type 3 (ALFPm3) gene, previously reported in P. monodon (Supungul et al., 2002) was found among the up-regulated genes but diversity due to amino acid changes was observed. Increase in ALFPm3 transcripts upon V. harveyi injection is in accordance with that found in the previous study.

Molecular Authentication and Genetic Polymorphism of Korean Ginseng (Panax ginseng C. A. Meyer) by Inter-Simple Sequence Repeats (ISSRs) Markers (ISSRs 마크에 의한 고려 인삼의 분자적 인증과 유전적 다형현상)

  • Bang, Kyong-Hwan;Lee, Sung-Woo;Hyun, Dong-Yun;Cho, Joon-Hyeong;Cha, Seon-Woo;Seong, Nak-Sul;Huh, Man-Kyu
    • Journal of Life Science
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    • v.14 no.3
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    • pp.425-428
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    • 2004
  • Molecular authentication and genetic polymorphism of Korean ginseng cultivars and accessions were investigated using ISSR (inter-simple sequence repeat amplification) markers. Five primers among 56 produced clear and reproducible DNA fragments among seven cultivars and accessions. A total of 43 bands ranging from 250 bp to 1,700 bp from five primers were scored. Average number of bands per primer was 8.6 and only nine bands were polymorphic across the six Panax ginseng from Korea. Especially Chunpoong cultivar exhibited the highest level of polymorphism, whereas other accessions did not showed almost any polymorphism. Consequently, these ISSR markers will be available to differentiate Chunpoong cultivar from other major Korean ginseng cultivars and accessions, such as Yunpoong, Hwangsukjong and Jakyungjong, at the DNA level.

DNA Fingerprinting by Amplified Fragment Length Polymorphism Markers in Rainbow Trout(Oncorhynchus mykiss)

  • Yoon, Jong-Man;Park, Sang-Hoon
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.559-560
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    • 2001
  • The objective of the present study was to analyze genetic variation and characteristics in rainbow trout(Oncorhynchus mykiss) using amplified fragment length polymorphism(AFLP) method as molecular genetic technique, to evaluate the usefulness of AFLP as genetic markers, and to compared the efficiency of agarose and polyacrylamide sequencing gels. The amplified products were performed by agarose and sequencing gel electrophoresis to detect AFLP band patterns, respectively. Using 9 primer combinations, total of 141 AFLP bands were produced, 108 bands(82.4%) of which were polymorphic in agarose gels. In sequencing gels, total of 288 bands were generated, and 220 bands (76.4%) were polymorphic. The level of bandsharing(BS) ranged from 0.18 to 0.32 for the 9 primer combinations tested, with a mean of 0.24. Consequently, AFLP markers of these rainbow trout could be used as genetic information such as species identification, genetic relationship or analysis of genome structure, and selection aids for genetic improvement of economically importment traits in fish species.

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Identification of antigenic proteins of lymphocystis disease virus (LCDV) by MALDI-TOF mass spectrometry

  • Chung, Chang-Kyun;Kim, Byung-Gwan;Jung, Myung-Hwa;Jung, Sung-Ju
    • Journal of fish pathology
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    • v.28 no.3
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    • pp.133-143
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    • 2015
  • The antigenic proteins of Lymphocystis disease virus (LCDV) from tumors of olive flounder, Paralichthys olivaceus, are described following characterization by mass spectrometry. In SDS-PAGE, predominant protein bands were observed at 114, 88, 70, 54, 52, 47, 42 and 24 kDa. Western blot analysis showed that antisera reacted strongly at molecular weights of 114, 67 and 54 kDa, and reacted weakly at molecular weights of 74, 70, 36, 24 and 22 kDa. In the identification of LCDV antigenic proteins by matrix-assisted laser desorption ionization (MALDI) TOF mass spectrometry, 10 of 14 excised bands consisted mostly of proteins with amino acid sequences that matched LCDV-C (lymphocystis disease virus isolate China) ORFs. Strong antigens with molecular weights of 114, 67 and 54 kDa were identified as LDVICp236 (chromosome segregation ATPase), LDVICp033 (membrane bound metallopeptidase) and LDVICp157 (hypothetical protein), respectively. Minor antigens with molecular weights of 70, 36, 24 and 22 kDa proteins were identified as LDVICp160 (acetyl-coA hydrolase), LDVICp213 (hypothetical protein), LDVICp039 (hypothetical protein) and LDVICp213 (hypothetical protein). However, the major capsid protein (LDVICp043) did not react with the polyclonal antibody.

Seed Purity Test and Genetic Diversity Evaluation Using RAPD Markers in Radish (Raphanus sativus L.)

  • Huh, Man-Kyu;Choi, Joo-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.4
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    • pp.346-350
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    • 2009
  • The cultivated radish (Raphanus sativus L.) is a major vegetable crop in the world wide and fast-growing species that grows inhabitats of six continents. It is very important to determine hybrid seed purity in the production of hybrid Brassica vegetable seeds to avoid unacceptable contamination with self-inbred (sib) seeds. The use of random amplified polymorphic DNA (RAPD) markers for evaluating seed purity in $F_2$-hybrid radish cultivars demonstrated. One hundred eighty seeds from the F1 male and female harvest were subsequently screened for seed purity using 13 primers. The 13 primers result in 17 cultivar-specific bands and 23 variable RAPD bands scored for cultivar. RAPD analysis of hybrid seeds from the harvest revealed 128 seeds tested except underdevelopment and decayed seeds were sibs. Especially, $F_2$ hybrids of radish, OPC13, OPD20 were presented clear hybrid bands. It maintains higher than average level of genetic diversity compared with their correspondent parents. RAPD amplification of DNA extracted from germinated individuals from the female harvest reveal that 10 of 208 seeds tested were self-inbred (4.8%). RAPD analysis of hybrid seeds from the male harvest revealed 7 of the 208 seeds tested were sibs (3.4%). The RAPD may lead to a better insight in to the hybrid seed purity.

Seed Purity Test and Evaluation in Isatis tinctoria var. yezoensis (Ohwi) Ohwi Using AFLP Markers (대청에서 AFLP를 이용한 종자순도검사와 평가)

  • Choi, Joo-Soo;Huh, Man-Kyu;Sung, Jung-Sook
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.3
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    • pp.198-203
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    • 2009
  • Isatis tinctoria var. yezoensis (Ohwi) Ohwi (Cruciferae) is one of major natural dyeing crops in the world and also have used as a medicinal plant in Korea. We evaluated seed purity in $F_1$-hybrid accessions using amplified fragment length polymorphism (AFLP) markers. One hundred sixty seeds from the male and female harvests were subsequently screened for seed purity with ten primers. The 13 accession-specific bands and many variable AFLP bands scored for accessions. Especially, E-AAC/M-CAA and E-AAG/M-CAT were presented clear hybrid bands for $F_1$ hybrids. $F_1$ hybrids maintained higher average level of genetic diversity compared with their correspondent parents. Self-inbred seeds from the female and male harvests were revealed 8.0% and 5.0%, respectively. The AFLP may lead to a better insight in to the hybrid seed purity test in I. tinctoria var. yezoensis.

Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.174-179
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    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

  • Khalilpour, Akbar;Santhanam, Amutha;Lee, Chun Wei;Saadatnia, Geita;Velusamy, Nagarajan;Osman, Sabariah;Mohamad, Ahmad Munir;Noordin, Rahmah
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1635-1642
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    • 2013
  • Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

The Comparison of Protein Patterns of Several Species in Bivalvia by SDS Polyacrylamide Gel Electrophoresis (SDS Polyacrylamide Gel 電氣泳動에 依한 斧足綱數種의 蛋白質패턴의 比較)

  • Park, Won-Chul;Ha, Man-Joon
    • The Korean Journal of Zoology
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    • v.28 no.2
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    • pp.61-70
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    • 1985
  • SDS-polyacrylamide gel electrophoresis for the proteins obtained from the plasma, Scapharca subcrenata and suapharca broughtonii, and for the proteins of muscles of several species in Bivalvia was performed. The protein patterns of plasma showed little difference between S. subcrenata and S. broughtonii in lower molecular weight weight proteins. However, the protein patterns of muscles of other species, which wre used in this study, were more shown in the lower molecular weights than the higher molecular weights in difference. Thus it is thought be an interesting fact. The protein band of blood corpuscles, 17,800 dalton, was not appeared in S. broughtonii, but this band was appeared in S. subcrenata. Henceforth this is the significantly important difference in these two species. But the protein patterns obtained from muscles of the species did not show a difference in a range of molecular weights between 10,000 and 100,000 daltons. Meanwhile, several protein bands obtained from Meretrix lusoria were similar to those of Mercenaria stimpsona. Hence, in this study, 6 protein bands which exist all species in Bivalvia and 4 characteristic protein bands in S. subcrenata and S. broughtoniionly, were investigated. And in four species of Eulamellibranchia, two protein bands in comon and the characteristic band of 23,000 dalton which is belong to Meretrix lusoria and Mercenaria stimdsona, were found. The molecular weights of the characteristic protein patterns, which are contained in each species, were measured and compared.

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NARROW-BAND SPECTROPHOTOMETRY OF COMET HALE-BOPP (C/1995 O1) NEAR PERIHELION I : PHOTOMETRIC BEHAVIOR OF $C_2$, $C_3$, CN MOLECULAR BANDS (헤일-밥 혜성(Comet Hale-Bopp, C/1995 O1)에 대한 근일점 근처에서의 분광측광 I : $C_2$, $C_3$, CN 분자 방출선의 측광학적 특성)

  • 성언창;김호일;윤재혁
    • Journal of Astronomy and Space Sciences
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    • v.17 no.2
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    • pp.199-210
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    • 2000
  • We present the results from narrow-band spectrophotometry of Comet Hale-Bopp (C/1995 O1) near perihelion obtained at Sobaeksan Optical Astronomy Observatory 61cm telescope equipped with PM 512 CCD camera(512$\times$512, 0.5"/pixel) and narrow-band filter set for the comet on 19 nights from February 21 to May 1, 1997. We discuss molecular emission band morphology and photometric behavior of Comet Hale-Bopp. The morphology of CN band shows more symmetric light distributions than $C_2$ or $C_3$ bands. On other hand, $C_2$ and $C_3$ band have more compact light distributions than CN band. Similar to wide-band image, molecular band morphology shows spiral structures at the core of the comet. The CN surface brightness variation with changing heliocentric distance shows difference from those of $C_2$ and $C_3$. The brightness, however, of these molecular bands near perihelion shows previously known 7day period light variations.

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