• Title/Summary/Keyword: Fragment size

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Expression Patterns of Bacillus subtilis Diacylglycerol Kinase Gene Induced by Physiological Stimuli (Bacillus subtilis dgk (diacylglycerol kinase) 유전자의 생리적 자극에 의한 유도발현)

  • Lee, Mi-Young;Suh, Seok-Jong;Lee, Jin-Hyung;Song, Bang-Ho;Kim, Jong-Cuk
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.15-20
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    • 2002
  • Diacylglycerol kinase (DGK) phosphorylates the second messenger diacylglycerol (DAG) to phosphatidic acid and it may play a role in signal transduction in Escherichia coli as well as in eukaryotic cells. In addition, DGK is important for microorganisms to adapt to several physiological stimuli. In Bacillus subtilis, the effect of stress on dgk transcription was examined by northern hybridization. The high level of dgk transcription was induced against high osmolarity, low pH value and low temperature. Transcriptional analysis revealed that the dgk gene and dgk upstream locus (ORF2, ORF3 and ORF4) were transcribed as a polycistronic mRNA to form an approximately 2.5 kb transcript.

Genetic Distances of Three White Clam (Meretrix lusoria) Populations Investigated by PCR Analysis

  • Kim, Dae-Hyun;Yoon, Jong-Man
    • Development and Reproduction
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    • v.18 no.2
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    • pp.89-98
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    • 2014
  • The twenty-one individuals of Meretrix lusoria were secured from Gunsan, Shinan and Yeonggwang on the coast of the Yellow Sea and the southern sea in the Korean Peninsula, respectively. Amplification of a single COI fragment (720 bp) was imagined, and no apparent size differences were observed in amplified fragments between Meretrix lusoria and M. petechialis individuals. The size of the DNA fragments also varied excitedly, from 200 to 1,600 bp. The oligonucleotides primer BION-08 produced the least loci (a total of 17), with an average of 2.43 in the Gunsan population, in comparison to the other primers used. Remarkably, the primer BION-13 detected 42 shared loci by the three populations, major and/or minor fragments of sizes 200 bp and 400 bp, respectively, which were identical in all samples. The dendrogram gained by the seven oligonucleotides primers highlight three genetic clusters: cluster 1 (GUNSAN 01 ~ GUNSAN 07), cluster 2 (SHINAN 08 ~ SHINAN 14) and cluster 3 (YEONGGWANG 15 ~ YEONGGWANG 21). The longest genetic distance among the twenty-one Meretrix lusoria individuals that displayed significant molecular differences was between individuals GUNSAN no. 01 and SHINAN no. 14 (genetic distance = 0.574). Comparatively, individuals of SHINAN population were fairly closely related to that of YEONGGWANG population. In this study, PCR analysis has discovered significant genetic distances between two white clam population pairs (P<0.05).

Association of Mahogany/Attractin Gene (ATRN) with Porcine Growth and Fat

  • Kim, Jong Bok;Zeng, Zhao Xun;Nam, Yu Jin;Kim, Younyoung;Yang, Shi Liu;Wu, Xiaolin;Lee, Chaeyoung
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1383-1386
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    • 2005
  • Associations of restriction fragment length polymorphism in porcine attractin (ATRN) gene with average daily gain (ADG), live weight (LWT) and back fat thickness (BFT) were studied with one exotic composition population (Xianghuang) and three local breeds (Ningxing, Shaziling and Daweizi) in China. Two types of alleles were detected; one with the band size of 700 bp (A) and the other with the band size of 450 and 250 bp (B), and their genotypes were classified as AA, AB and BB. Polymorphisms were observed in the exotic breed, but not in the three local breeds. Individuals with the genotype BB showed larger ADG and LWT than those with AA in Xianghuang population (p<0.05). The breed specific effects of the porcine attractin gene in this study implied that identification of the polymorphisms and determination of genetic effects on phenotypes should be performed in the target populations.

Isolation and Partial Characterization of Bacteriophage from Oyster (생굴로부터 bacteriophage의 분리 및 부분특성)

  • 김영희
    • Journal of Environmental Science International
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    • v.5 no.5
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    • pp.605-610
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    • 1996
  • The bacteriophage from the fresh oyster, Crassostrea Virginica which is specific to the marine bacterium was isolated and characterized. Among the foci different vibrio species and the five different serotypes of Vibrio parahaemolyticus host strains tested, only two strains of the parahaemolyticus possessing K17, K52 antigens were highly sensitive to the phage. The size of the isolated plaque was 0.4mm and the electron microscopic head size of the isolated phage was about 67 nm long and 83 nm wide. PFU/ml was 1.25$\times$ $10^{11}$. The phase was sensitive to chloroform but resistant to acetone or methanol. The assay of the isolated phase nucleic acid was deoxyribonucleic acid. The restriction enzyme pattern showed 14 fragment from Hind III and 4 fragments from Eco R I. Two different antigenic groups showed-similar restriction enzyme patterns.

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Prediction of rock fragmentation and design of blasting pattern based on 3-D spatial distribution of rock factor

  • Sim, Hyeon-Jin;Han, Chang-Yeon;Nam, Hyeon-U
    • 지반과기술
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    • v.3 no.3
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    • pp.15-22
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    • 2006
  • The optimum blasting pattern to excavate a quarry efficiently and economically can be determined based on the minimum production cost, which is generally estimated according to rock fragmentation. Therefore, it is a critical problem to predict fragment size distribution of blasted rocks over an entire quarry. By comparing various prediction models, it can be ascertained that the result obtained from Kuz-Ram model relatively coincides with that of field measurements. Kuz-Ram model uses the concept of rock factor to signify conditions of rock mass such as block size, rock jointing, strength and others. For the evaluation of total production cost, it is imperative to estimate 3-D spatial distribution of rock factor for the entire quarry. In this study, a sequential indicator simulation technique is adopted for estimation of spatial distribution of rock factor due to its higher reproducibility of spatial variability and distribution models than Kriging methods. Further, this can reduce the uncertainty of predictor using distribution information of sample data. The entire quarry is classified into three types of rock mass and optimum blasting pattern is proposed for each type based on 3-D spatial distribution of rock factor. In addition, plane maps of rock factor distribution for each ground level are provided to estimate production costs for each process and to make a plan for an optimum blasting pattern.

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Detection of Pectobacterium chrysanthemi Using Specific PCR Primers Designed from the 16S-23S rRNA Intergenic Spacer Region

  • Kwon, Soon-Wo;Myung, In-Sik;Go, Seung-Joo
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.252-256
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    • 2000
  • The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

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Frequencies, Inheritance of Porcine FSH-${\beta}$ Retroposon and its Association with Reproductive Traits

  • Li, Feng'e;Xiong, Yuanzhu;Deng, Changyan;Jiang, Siwen;Zheng, Rong
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.2
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    • pp.179-183
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    • 2002
  • The fragment in intron I of FSH-${\beta}$ gene was amplified by PCR. According to the polymorphism, we analyzed the distribution of FSH-${\beta}$ retroposon in different pig breeds; its inheritance pattern in Large White${\times}$Meishan reference family; and the association of FSH-${\beta}$ retroposon with litter size, female reproductive organs measurement, ultrasonic backfat and other traits. The results showed that almost each Chinese indigenous pig had the retroposon, while foreign pig breeds rarely had; the frequencies of porcine FSH-${\beta}$ retroposon were strongly associated with breeds (p<0.01); the pattern of inheritance was consistent with Mendelian fashion; total number born (TNB) and number born alive (NBA) were increased per FSH-${\beta}$ retroposon (p<0.01) with additive effects of 1.2-1.8 and 1.4-1.8 pigs/litter, respectively; between the FSH-${\beta}$ retroposon carriers and non-carriers, there was an insignificant difference in the measurement of female reproductive organs, body weight at birth, backfat thickness, loin meat height, lean meat percentage, teat number, days to 100 kg, and average daily gain.

Complementary DNA Cloning of Genomic RNA in Orchid Strain of Tobacco Mosaic Virus

  • Won Mok Park
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.349-355
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    • 1994
  • Viral RNA was extracted from a purified orchid strain of tobacco mosaic virus (TMV-O) from Cymbidium "Grace Kelly". Polyadenylated viral RNAs were primed with Not I-oligo (dT) primer-adapter. First-strand cDNAs were reversely transcribed by Moloney murine leukaemia virus reverse transcriptase (RNAse H-), and then second-strand cDNAs were synthesized by RNase H and DNA polymerase I. The resulting double-stranded cDNAs were ligated into pSPORT1 vector and transformed into competent E. coli strain JM109 cells. The size of cDNAs within the recombinant plasmids was ranging from 0.9 to 3.9 kb. Among the selected clones, pTMO-0205 and -0210 covered the 3' half and the 5' half of the viral genomic RNA, respectively, which were covering more than 99% of the viral genemo size based on sequencing analysis. Two cDNA fragments which were 3.1 kb BamHI and NotI fragement released from pTMO-0.205 and 3.3 kb SalI and BamHI fragment released from pTMO-0210 were ligated with T4 DNA ligase. The clone was almost entire length, lacking only 31 nucleotides from the 5' terminus based on the sequencing result. This method was shown to be efficiently applicable to other plant viral gnomic RNA for the construction of cDNA.n of cDNA.

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Detection of Fish Virus by Using Immunomagnetic Separation and Polymerase Chain Reaction (IMS-PCR)

  • KIM Soo Jin;OH Hae Keun;CHOI Tae-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.6
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    • pp.948-955
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    • 1997
  • Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

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Genetic Diversity of Korean Cyanobacteria determined by DNA polymorphisms within the Phycocyanin Locus (Phycocyanin locus내의 DNA Polymorphism에 의한 한국산 Cyanobacteria의 유전적 다양성)

  • 박진숙;권주리;유순애
    • Korean Journal of Microbiology
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    • v.36 no.4
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    • pp.249-253
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    • 2000
  • The genetic diversity among Korean cyanobacteria was assessed by restriction fragment length polymorphism(RFLP) analysis of PCR products from the phycocyanin locus. Strains of all the genera tested were successfully amplified, and the size of amplified fragments was approximately 700bp. The restriction patterns generated by AluI, MspI, and HaeIII were conserved for strains within each of genera studied and were specific to the genus level. Intrageneric delineation of strains was revealed by the enzyme, CfoI for members of genera Anabeana and Synechocystis. Phenogram derived from the different RFLP patterns revealed a coherent cluster among Anabeana, Chlorogloea, and Synechocystis strains. PC-RFLP methods provided useful tools for classification of cyanobacteria.

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