• Title/Summary/Keyword: sequence length

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Development of PCR-based markers for discriminating Solanum berthaultii using its complete chloroplast genome sequence

  • Kim, Soojung;Cho, Kwang-Soo;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.207-216
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    • 2018
  • Solanum berthaultii is one of the wild diploid Solanum species, which is an excellent resource in potato breeding owing to its resistance to several important pathogens. On the other hand, sexual hybridization between S. berthaultii and S. tuberosum (potato) is limited because of their sexual incompatibility. Therefore, cell fusion can be used to introgress various novel traits from this wild species into the cultivated potatoes. After cell fusion, it is crucial to identify fusion products with the aid of molecular markers. In this study, the chloroplast genome sequence of S. berthaultii obtained by next-generation sequencing technology was described and compared with those of five other Solanum species to develop S. berthaultii specific markers. A total sequence length of the chloroplast genome is 155,533 bp. The structural organization of the chloroplast genome is similar to those of the five other Solanum species. Phylogenic analysis with 25 other Solanaceae species revealed that S. berthaultii is most closely located with S. tuberosum. Additional comparison of the chloroplast genome sequence with those of the five Solanum species revealed 25 SNPs specific to S. berthaultii. Based on these SNPs, six PCR-based markers for differentiating S. berthaultii from other Solanum species were developed. These markers will facilitate the selection of fusion products and accelerate potato breeding using S. berthaultii.

Cloning and Sequencing of Nonstructural Protein (NSP4) Gene of Bovine Rotavirus Isolated in Korea (국내분리 소 로타바이러스 NSP4 유전자의 크로닝 및 염기서열 분석)

  • Back, Myoung-Soon;Kim, Won-Yong;Kang, Shien-Young
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.1-10
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    • 2000
  • The nonstructural protein NSP4, encoded by gene 10 of rotavirus, has been shown to playa role in viral assembly and known to be an enterotoxin, causing diarrhea in mouse pups. NSP4 gene was cloned from CBNU-2 (virulent bovine rotavirus/diarrheic fecal sample) and CBNU-1 (cell-culture adapted bovine rotavirus/isolated from CBNU-2 and 75 times passaged on MA104 cells), respectively, by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced and compared. The sequence data indicated that the NSP4 genes of bovine rotavirus (BRV) were 751 bases in length and encoded one open reading frame of 175 amino acids beginning at base 42 and terminating at base 569. Differences in nucleotide sequence between CBNU-2 and CBNU-1 were observed at 6 positions (base 274, 296, 391, 394, 396 and 579). NSP4 gene of BRV exhibited a high degree of nucleotide (90% and 94%) and amino acid sequence (91% and 97%) homology with those of SA11 and UK but a low degree of nucleotide (77% and 79%) and amino acids sequence (81% and 85%) homology with those of Wa and OSU.

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Effect of random Shine-Dalarno sequence on the expression of Bovine Growth Hormone Gene in Escherichia coli (대장균에서 무작위 샤인-달가노 서열이 소성장호르몬 유전자 발현에 미치는 영향)

  • 나경수;나경수;백형석;이용세
    • Journal of Life Science
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    • v.10 no.4
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    • pp.422-430
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    • 2000
  • In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

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Identification and Characterization of New Copia-like Retrotransposon Osr1 in Rice

  • Lee, Yong-Hwan;Jwa, Nam-Soo;Park, Sook-Young;Park, Chan-Ho;Han, Seong-Sook
    • The Plant Pathology Journal
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    • v.19 no.1
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    • pp.57-63
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    • 2003
  • An insertion sequence identified as a solo long terminal repeat (LTR) of a new rice copia-like retrotransposon was detected in the ORE of the Pi-b gene from the rice cv. Nipponbare, and was designated as Osr1. Osr1 consists of a 6386 bp nucleotide sequence including 965 bp LTRs on both ends with an 82% nucleotide sequence identity to the wheat Tarl retrotransposon on reverse transcriptase. Nucleotide divergence was noted among the individual LTRs, as well as the coding region of Osr1. Various restriction fragment length polymorphism (RFLP) of LTR were detected in indica cultivars, whereas, only a few could be detected in the japonica cultivars. The population of Osr1 is lower in the wild-type rice compared with that in the domesticated cultivars. The insertion of LTR sequence in the h-b gene in the susceptible cultivar suggested that retro-tyansposon-mediated insertional mutation might play an important role in the resistance breakdown, as well as in the evolution of resistance genes in rice.

Genetic Studies on Korean Anurans: Length and Restriction Site Variation in the Mitochondrial DNA of Tree Frogs, Hyla japonica and H. suweonensis (한국산 무미류에 대한 유전학적 연구 : 청개구리속 2종(Hyla japonira, H. suweonensis)에 대한 mtDNA의 크기 및 제한효소 인식위치의 변이)

  • 이혜영;박창신
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.219-225
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    • 1992
  • The genetic variation in mitochondrial DNA (mtDNA) was analysed within and between two species of tree frogs. Hyla japonica and H. suweonensis from South Korea. Purified mtDNAs were digested with each of 11 restriction enLvmes which cleave at six base recognition sequences. The genome size of H. iaponica revealed ho types (20.0 $\pm$ 0.3 and 19.6 $\pm$ 0.3 kb) and this difference is explained by either addition or deletion of about 0.4 kb fragment. On the other hand, the genome sire of H. suueonensis was about 19.0 $\pm$ 0.4 kb only. For the analysis, level of fragment homology (F) and nucleotide sequence divergence (p) were estimated from comparisons of digestion profiles. Among four populations of H. iaponica, substantial mean sequence divergence was 0.017 (range 0.001-0.026); between identical types, 0.001 IslilaRl type) and 0.004 (Large type) respectively; between different ones, 0.024 (range 0.023-0.026). The level of sequence divergence between he species was 0.142 (range 0.131-0.146). This result suggested that he species ㅂwere distinctly differentiated species. The divergence time between ko species was estimated 7.1 million years.

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Comparison between of the Attenuated BR-Oka and the Wild Type Strain of Varicella Zoster Virus (VZV) on the DNA level

  • Lim, Sang-Min;Song, Seong-Won;Kim, Sang-Lin;Jang, Yoon-Jung;Kim, Ki-Ho;Kim, Hong-Jin
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.418-423
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    • 2000
  • Oka strain VR-795 (Varicella Zoster Virus, VZV) of American Type Culture Collection (ATCC) has been used for chickenpox vaccine production. In order to use this strain for vaccine production, the strain must be identified and its stability must be confirmed. The identification of the Oka strain has been confirmed using Restriction Fragment Length Polymorphism (RFLP) and DNA sequence analysis of glycoprotein-II (gp-II). The amino acid sequences of Oka deduced from the DNA sequence of gp-II have changed at three amino acids against Ellen and at one amino acid against Webster. To prove the stability of the Oka strain during the passage, RFLP and DNA sequence analyses were also used with 11, 15 and 23 times of virus passage. We found that the Oka strain was stable at passages of up to 23 times, based on the RFLP and DNA sequence analyses. The confirmed Oka strain was renamed as BR-Oka for the purposes of chickenpox vaccine production.

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Cloning and Sequence Analysis of Ribosomal Protein S4 cDNA from Root of Panax ginseng

  • In Jun-Gyo;Lee Bum-Soo;Song Won-Seob;Bae Chang-Hyu;Choi Seong-Kyu;Yang Deok-Chun
    • Plant Resources
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    • v.8 no.2
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    • pp.110-115
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    • 2005
  • Ribosomal protein complex with ribosomal RNA to form the subunits of the ribosome serve essential functions in protein synthesis. A full-length cDNA (PRPS4) encoding ribosomal protein S4 has been isolated and its nucleotide sequence determined in ginseng plant (Panax ginseng). A PRPS4 cDNA is 1105 nucleotides long and has an open reading frame of 792 bp with a deduced amino acid sequence of 264 residues (pI 10.67). The deduced amino acid sequence of PRPS4 matched to the previously reported ribosomal protein S4 genes. Their degree of amino acid identity ranged from 68 to $92\%$. Phylogenetic analysis based on the amino acid residues showed that the PRPS4 grouped with ribosomal protein S4 of S. tuberosum (CAA54095).

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Genetic Variation of Rice Populations Estimated Using nrDNA ITS Region Sequence

  • Wang, Dong;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.249-255
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    • 2014
  • The rice belonging to Oryza sativa is not only has significant economic importance, for it is the major source of nutrition for about 3 billion all around the world. But also plays a vital role as a model organism, because it has a number of advantages to be a model plant, such as efficient transformation system and small genome size. Many methods and techniques have been conducted to attempt to distinguish different Oryza sativa species, such as amplified fragment length polymorphism (AFLP), random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and so on. However, studies using sequence analysis of internal transcribed spacer (ITS), a region of ribosomal RNA has not been reported until now. This study was undertaken with an aim to understand the phylogenetic relationships among sixteen isolates of Oryza sativa collected from abroad and fifteen isolates collected from Korea, using ribosomal RNA (rRNA) internal transcribed spacer (ITS) sequences to compare the phylogeny relationships among different Oryza sativa species. The size variation obtained among sequenced nuclear ribosomal DNA (nrDNA) ITS region ranged from 515bp to 1000bp. The highest interspecific genetic distance (GD) was found between Sfejare 45 (FR12) and Anapuruna (FR15). Taebong isolate showed the least dissimilarity of the ITS region sequence with other thirty isolates. This consequence will help us further understanding molecular diversification in intra-species population and their phylogenetic analysis.

Genetic Clarification of Auricularia heimuer Strains Bred and Cultivated in Korea Using the ITS and IGS1 rDNA Region Sequences

  • Nitesh Pant;HyeongJin Noh;Won-Ho Lee;Seong Hwan Kim
    • Mycobiology
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    • v.51 no.2
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    • pp.109-113
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    • 2023
  • Auricularia is one of the broadly cultivated edible mushrooms in Korea. Most of the Korean Auricularia strains used for cultivation and breeding are known as A. auricula-judae. Recently, this species has been reported to belong to a species complex. Therefore, this study was carried out to genetically clarify the bred and cultivated Korean A. auricula-judae strains. The internal transcribed spacer (ITS) and IGS1 rDNA region sequences were determined from 10 A. auricula-judae strains by PCR and sequencing. Variation in the nucleotide sequence and sequence length of the two rDNA regions were found among the seven A. auricula-judae strains. A maximum-likelihood (ML) phylogenetic tree based on the ITS sequences clearly placed all the 10 Korean A. auricula-judae strains in the A. heimuer clade of the A. auriculajudae complex. A. heimuer is diverged from A. auricula-judae. An ML phylogenetic tree based on the IGS1 sequences revealed the close relationship between Korean A. heimuer strains to Chinese A. heimuer strains. But each strain could be distinguishable by the IGS1 sequence. Furthermore, progeny strains in the seven Korean strains could be differentiated from their parental strains by the IGS1 sequence based phylogenetic tree. Our results are expected to be used to complement the distinction of domestic Auricularia cultivars.

Effects of the mascara and eye shadow on theMR image distortion (자기공명영상 왜곡에서 마스카라와 아이섀도의 영향)

  • Lee, Hyun-Yong;Shin, Oun-Jae;Park, Byung-Rae
    • Journal of radiological science and technology
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    • v.28 no.1
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    • pp.25-32
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    • 2005
  • Purpose : To evaluate the degree of the artifact which is caused by the mascara and the eye shadow when acquiring MR images and compare the difference of the image distortion according to the change of various pulse sequence. Material and Method : The popular domestic mascara and eye shadow products were selected from three different companies respectively and divided into two groups mascara (M1, M2, M3 ), eye shadow (E1, E2, E3). Self-designed quadrature type saddle coil which has 4 cm inside diameter, 8 cm length and which is for both Tx and Rx was used. MR image was acquired respectively after applying the mascara to the tape from study 1, the eye shadow to the tape from study 2 and adding the eye shadow to the mascara from study 3. The FSE(fast spin echo), the SE(spin echo), the GE(gradient echo) were used as pulse sequences. The degree of the image distortion which was measured from each sequence was analyzed in quality and quantity. Result : The mascara and the eye shadow caused the artifacts to the MR images partially and induced the image distortion. There was a little difference in terms of the degree of artifact according to the change of pulse sequence. From the study 3 in which the eye shadow was applied to the mascara, on the axial plane image, the width of artifact was 16.73 mm in the GE pulse sequence, 6.64 mm in the SE pulse sequence, and 6.19 mm in the FSE pulse sequence. The degree of the artifact appeared highly in order of the GE, the SE and the FSE. On the sagittal plane image, the length of artifact was 22.84 mm in the GE, 17.81 mm in the SE and it appeared highly with the SE and the FSE technique order. Conclusion : When examining the eyeball and the brain of a woman with the mascara and the eye shadow, we have to consider that the artifact caused by them can have an effect on the image diagnosis. We concluded that it is more suitable for a brain and a eyeball T2 emphasizing image to use the FSE technique than the GE technique.

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