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Development of SSLP Marker Targeted to P34 Null Gene in Soybean (콩 P34 단백질 결핍 유전자를 이용한 SSLP 마커 개발)

  • Yang, Kiwoung;Ko, Jong-Min;Lee, Young-Hoon;Jeon, Myeong Gi;Jung, Chan-Sik;Baek, In-Youl;Kim, Hyun-Tae;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.502-506
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    • 2010
  • Soybean seed possesses about 15 allergenic proteins recognized by IgEs from soy-sensitive human. The allergenic impact of soybean proteins limit its extensive usage in a broad range of processed foods. Soybean protein P34 or Gly m Bd 30k of the cysteine protease family is one of the major allergen of the soybean seed. P34-null soybean, PI567476, was identified among soybean (Glycine max & Glycine soja Sieb. and Zucc) of approximately 16,226 accessions from USDA soybean germplasm screened. Also, for P34 gene (Williams 82; whole genome sequence cultivar) and P34 null gene (PI567476) comparative analysis of sequences listed in the NCBI database showed the presence of a SSLP (Simple Sequence Length Polymorphism) of 4 base pair. So, a SSLP marker was designed to reveal the polymorphism of the locus. In this study, a population of 339 $F_2$ recombinant inbred lines generated by cross between Taekwang (Glycine max) and PI567476 was used to select $F_{2:3}$ plant of a P34 null gene. The result separation rate Taekwang type, heterozygous type and PI567476 type were shown in 85: 187: 67 since single gene is concerned in as the separation rate of 1:2:1 in $X^2{_{0.05}}=5.99$, df=2. In future, selected plant will identify protein level, whether P34 null protein is equal to P34 null gene.

Linearly Polarized 1-kW 20/400-㎛ Yb-doped Fiber Laser with 10-GHz Linewidth (선편광된 10 GHz 선폭의 1 kW급 20/400-㎛ 이터븀 첨가 광섬유 레이저)

  • Jung, Yeji;Jung, Minwan;Lee, Kangin;Kim, Taewoo;Kim, Jae-Ihn;Lee, Yongsoo;Cho, Joonyong
    • Korean Journal of Optics and Photonics
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    • v.32 no.3
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    • pp.120-125
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    • 2021
  • We have developed a linearly polarized high-power Yb-doped fiber laser in the master oscillator power amplifier (MOPA) scheme for efficient spectral beam combining. We modulated the phase of the seed laser by pseudo-random binary sequence (PRBS), with the bit length optimized to suppress stimulated Brillouin scattering (SBS), and subsequently amplified seed power in a 3-stage amplifier system. We have constructed by coiling the polarization-maintaining (PM) Yb-doped fiber, with core and cladding diameters of 20 ㎛ and 400 ㎛ respectively, to a diameter of 9-12 cm for suppression of the mode instability (MI). Finally, we obtained an output power of 1.004 kW with a slope efficiency of 83.7% in the main amplification stage. The beam quality factor M2 and the polarization extinction ratio (PER) were measured to be 1.12 and 21.5 dB respectively. Furthermore, the peak-intensity difference between the Rayleigh signal and SBS signal was observed to be 2.36 dB in the backward spectra, indicating that SBS is successfully suppressed. In addition, it can be expected that the MI does not occur because not only there is no decrease in slope efficiency, but also the beam quality for each amplified output is maintained.

The Complete Mitochondrial Genome and Molecular Phylogeny of the Flathead Platycephalus cultellatus Richardson, 1846 from Vietnam (Teleostei; Scorpaeniformes) (베트남 Platycephalus cultellatus Richardson, 1846 (Teleostei; Scorpaeniformes)의 전장 미토콘드리아 유전체와 분자계통)

  • Tran, Biet Thanh;Nguyen, Tu Van;Choi, Youn Hee;Kim, Keun-Yong;Heo, Jung Soo;Kim, Keun-Sik;Ryu, Jung-Hwa;Kim, Kyeong Mi;Yoon, Moongeun
    • Korean Journal of Ichthyology
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    • v.33 no.4
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    • pp.217-225
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    • 2021
  • The family Platycephalidae is a taxonomic group of economically important demersal flathead fishes that predominantly occupy tropical or temperate estuaries and coastal environments of the Indo-Pacific oceans and the Mediterranean Sea. In this study, we for the first time analyzed the complete mitochondrial genome (mitogenome) of the flathead Platycephalus cultellatus Richardson, 1846 from Vietnam by Next Generation Sequencing method. Its mitogenome was 16,641 bp in total length, comprising 13 protein-coding genes (PCGs), two ribosomal RNA genes, and 22 transfer RNA genes. The gene composition and order of the mitogenome were identical to those of typical vertebrates. The phylogenetic trees were reconstructed based on the concatenated nucleotide sequence matrix of 13 PCGs and the partial sequence of a DNA barcoding marker, cox1 in order to determine its molecular phylogenetic position among the order Scorpaeniformes. The phylogenetic result revealed that P. cultellatus formed a monophyletic group with species belonging to the same family and consistently clustered with one nominal species, P. indicus, and two Platycephalus sp. specimens. Besides, the cox1 tree confirmed the taxonomic validity of our specimen by forming a monophyletic clade with its conspecific specimens. The mitogenome of P. cultellatus analyzed in this study will contribute valuable information for further study on taxonomy and phylogeny of flatheads.

PCR-based markers to select plastid genotypes of Solanum acaule (Solanum acaule 색소체 유전자형 선발을 위한 특이적 분자마커 개발)

  • Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.178-186
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    • 2022
  • The tetraploid Solanum acaule is a wild potato species from Bolivia widely used for potato breeding because of its diverse attractive traits, including resistance to frost, late blight, potato virus X, potato virus Y, potato leafroll virus, potato spindle tuber viroid, and cyst nematode. However, the introgression of useful traits into cultivated potatoes via crossing has been limited by differences in endosperm balance number between species. Somatic fusion could be used to overcome sexual reproduction barriers and the development of molecular markers is essential to select proper fusion products. The chloroplast genome of S. acaule was sequenced using next-generation sequencing technology and specific markers for S. acaule were developed by comparing the obtained sequence with those of seven other Solanum species. The total length of the chloroplast genome is 155,570 bp, and 158 genes were annotated. Structure and gene content were very similar to other Solanum species and maximum likelihood phylogenetic analysis with 12 other species belonging to the Solanaceae family revealed that S. acaule is very closely related to other Solanum species. Sequence alignment with the chloroplast genome of seven other Solanum species revealed four InDels and 79 SNPs specific to S. acaule. Based on these InDel and SNP regions, one SCAR marker and one CAPS marker were developed to discriminate S. acaule from other Solanum species. These results will aid in exploring evolutionary aspects of Solanum species and accelerating potato breeding using S. acaule.

Molecular Characterization of an Isolate of Bean Common Mosaic Virus First Identified in Gardenia Using Metatranscriptome and Small RNA Sequencing

  • Zhong-Tian Xu;Hai-Tao Weng;Jian-Ping Chen;Chuan-Xi Zhang;Jun-Min Li;Yi-Yuan Li
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.73-82
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    • 2024
  • Gardenia (Gardenia jasminoides) is a popular and economically vital plant known for its ornamental and medicinal properties. Despite its widespread cultivation, there has been no documentation of plant viruses on gardenia yet. In the present study, gardenia leaves exhibiting symptoms of plant viral diseases were sampled and sequenced by both metatranscriptome and small RNA sequencing. As a consequence, bean common mosaic virus (BCMV) was identified in gardenia for the first time and named BCMV-gardenia. The full genome sequence of BCMV-gardenia is 10,054 nucleotides (nt) in length (excluding the poly (A) at the 3' termini), encoding a large polyprotein of 3,222 amino acids. Sequence analysis showed that the N-termini of the polyprotein encoded by BCMV-gardenia is less conserved when compared to other BCMV isolates, whereas the C-termini is the most conserved. Maximum likelihood phylogenetic analysis showed that BCMVgardenia was clustered closely with other BCMV isolates identified outside the leguminous plants. Our results indicated that the majority of BCMV-gardenia virus-derived small interfering RNAs (vsiRNAs) were 21 nt and 22 nt, with 21 nt being more abundant. The first nucleotide at the 5' termini of vsiRNAs derived from BCMV-gardenia preferred U and A. The ratio of vsiRNAs derived from sense (51.1%) and antisense (48.9%) strands is approaching, and the distribution of vsiRNAs along the viral genome is generally even, with some hot spots forming in local regions. Our findings could provide new insights into the diversity, evolution, and host expansion of BCMV and contribute to the prevention and treatment of this virus.

Development of PCR-based markers for selecting plastid genotypes of Solanum hjertingii (Solanum hjertingii 색소체 유전자형 선발을 위한 PCR 기반 분자마커 개발)

  • Tae-Ho Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.34-44
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    • 2023
  • The tetraploid Solanum hjertingii, a wild tuber-bearing species from Mexico is a relative of potato, S. tuberosum. The species has been identified as a potential source of resistance to blackening for potato breeding. It does not exhibit enzymatic browning nor blackspot which are physiological disorders. However, due to their sexual incompatibility, somatic hybridization between S. hjertingii and S. tuberosum must be used to introduce various traits from this wild species into potato. After somatic hybridization, molecular markers are essential for selecting fusion products. In this study, the chloroplast genome of S. hjertingii was sequenced by next-generation sequencing technology and compared with those of other Solanum species to develop specific markers for S. hjertingii. The chloroplast genome has a total sequence length of 155,545 bp, and its size, gene content, order and orientation are similar to those of the other Solanum species. Phylogenic analysis including 15 other Solanaceae species grouped S. hjertingii with S. demissum, S. hougasii, and S. stoloniferum. After detailed comparisons of the chloroplast genome sequence with eight other Solanum species, we identified one InDel and seven SNPs specific to S. hjertingii. Based on these, five PCR-based markers were developed for discriminating S. hjertingii from other Solanum species. The results obtained in this study will aid in exploring the evolutionary aspects of Solanum species and accelerating breeding using S. hjertingii.

Molecular Cloning and Characterization of a New cDNA Encoding Hyoscyamine 6β-hydroxylase from Roots of Anisodus acutangulus

  • Kai, Guoyin;Chen, Junfeng;Li, Li;Zhou, Genyu;Zhou, Limin;Zhang, Lei;Chen, Yuhui;Zhao, Linxia
    • BMB Reports
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    • v.40 no.5
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    • pp.715-722
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    • 2007
  • A new full-length cDNA encoding hyoscyamine $6\beta$-hydroxylase (designated as aah6h, GenBank Accession No. EF187826), which catalyzes the last committed step in the scopolamine biosynthetic pathway, was isolated from young roots of Anisodus acutangulus by rapid amplification of cDNA ends (RACE) for the first time. The full-length cDNA of aah6h was 1380 bp and contained a 1035 bp open reading frame (ORF) encoding a deduced protein of 344 amino acid residues. The deduced protein had an isoelectric point (pI) of 5.09 and a calculated molecular mass of about 38.7 kDa. Sequence analyses showed that AaH6H had high homology with other H6Hs isolated from some scopolamine-producing plants such as Hyoscyamus niger, Datura metel and Atropa belladonna etc. Bioinformatics analyses results indicated AaH6H belongs to 2-oxoglutarate-dependent dioxygenase superfamily. Phylogenetic tree analysis showed that AaH6H had closest relationship with H6H from A. tanguticus. Southern hybridization analysis of the genomic DNA revealed that aah6h belonged to a multi-copy gene family. Tissue expression pattern analysis firstly founded that aah6h expressed in all the tested tissues including roots, stems and leaves and indicated that aah6h was a constitutive-expression gene, which was the first reported tissue-independent h6h gene compared to other known h6h genes.

Screening and Characterization of LTR Retrotransposons in the genomic DNA of Pleurotus eryngii (큰느타리버섯 유전체내 LTR Retrotransposon 유전자 탐색 및 특성연구)

  • Kim, Sinil;Le, Quy Vang;Kim, Sun-Mi;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.50-56
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    • 2014
  • Transposable elements (TEs) are mobile DNA elements that often cause mutations in genes and alterations in the chromosome structure. In order to identify and characterize transposable elements (TEs) in Pleurotus eryngii, a TE-enriched library was constructed using two sets of TE-specific degenerated primers, which target conserved sequences of RT and RVE domains in fungal LTR retrotransposons. A total of 256 clones were randomly chosen from the library and their insert sequences were determined. Comparative investigation of the insert sequences with those in repeat element database, Repbase, revealed that 71 of them were found to be TE-related fragments with significant similarity to LTR retrotransposons from other species. Among the TE sequences, the 70 TEs were Gypsy-type LTR retrotransposons, including 20 of MarY1 from Tricholoma matsutake, 26 of Gypsy-8_SLL from Serpula lacrymans, and 16 of RMER17D_MM from mouse, whereas a single sequence, Copia-48-PTR, was found as only Copia-type LTR retrotransposon. Southern blot analysis of the HindIII-digested P. eryngii genomic DNA showed that the retrotransposon sequences similar to MarY1 and Gypsy-8_SLL were contained as high as 14 and 18 copies per genome, respectively, whereas other retrotransposons were remained low. Moreover, both of the two Gypsy retrotransposons were expressed in full length mRNA as shown by Northern blot analysis, suggesting that they were functionally active retrotransposons.

Characterization of Phosphoinositide-3-kinase, Class 3 (PIK3C3) Gene and Association Tests with Quantitative Traits in Pigs

  • Kim, J.H.;Choi, B.H.;Lim, H.T.;Park, E.W.;Lee, S.H.;Seo, B.Y.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1701-1707
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    • 2005
  • This study deals with the characterization of porcine PIK3C3 and association tests with quantitative traits. PIK3C3 belongs to the class 3 PI3Ks that participate in the regulation of hepatic glucose output, glycogen synthase, and antilipolysis in typical insulin target cells such as those in the such as liver, muscle system, and fat. On the analysis of full-length mRNA sequence, the length of the PIK3C3 CDS was recorded as 2,664 bps. As well, nucleotide and amino acid identities between human and pig subjects were 92% and 99%, respectively. Five SNPs were detected over 5 exons. We performed genotyping by using a SNP C2604T on exon24 for 145 F$_2$ animals (from a cross between Korean native boars and Landrace sows) by PCR-RFLP analysis with Hpy8I used to investigate the relationship between growth and fat depot traits. In the total association analysis, which doesn' consider transmission disequilibrium, the SNP showed a significant effect (p<0.05) on body weight and carcass fat at 30 weeks of age as well as a highly significant effect (p<0.01) on back fat. In an additional sib-pair analysis, C allele still showed positive and significant effects (p<0.05) on back fat thickness and carcass fat. Moreover, the effects of C allele on the means of within-family components for carcass fat and back fat were estimated as 2.76 kg and 5.07 mm, respectively. As a result, the SNP of porcine PIK3C3 discovered in this study could be utilized as a possible genetic marker for the selection of pigs that possess low levels of back fat and carcass fat at the slaughter weight.

Identification and characteristics of DDX3 gene in the earthworm, Perionyx excavatus (팔딱이 지렁이(Perionyx excavatus) DDX3 유전자의 동정 및 특성)

  • Park, Sang Gil;Bae, Yoon-Hwan;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.23 no.1
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    • pp.70-81
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    • 2015
  • Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.