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Chrysanthemum Chlorotic Mottle Viroid-Mediated Trafficking of Foreign mRNA into Chloroplasts

  • Baek, Eseul;Park, Minju;Yoon, Ju-Yeon;Palukaitis, Peter
    • Research in Plant Disease
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    • v.23 no.3
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    • pp.288-293
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    • 2017
  • Chrysanthemum chlorotic mottle viroid (CChMVd) fused to the leader sequence of a reporter gene (mRFP) expressed transiently in agroinfiltrated Nicotiana benthamiana, was used to show that CChMVd can traffic into chloroplasts, thought to be the site of its replication. Fluorescence from mRFP was detected in chloroplasts, but only if the viroid transcription fusions were present, either from the full-length 400-nt CChMVd, or each of two partial fragments (nucleotides 125 to 2 and 231 to 372). The mRFP and its mRNA were detected by western blotting and RT-PCR, respectively, in tissue extracts of plants infiltrated by each fusion construct. Isolated chloroplasts were shown by RT-PCR to contain the RNA sequences of both CChMVd and mRFP, if both were present, but not the mRFP sequence in the absence of the viroid sequences. The results suggest that RNA trafficking was probably due to an RNA structure, and not a particular sequence, as discussed.

Synthesis of 90/102(170)/150 linear CA using 90/150 linear CA (90/150 선형 CA를 이용한 90/102(170)/150 선형 CA 합성)

  • Choi, Un-Sook;Cho, Sung-Jin;Kim, Han-Doo;Kwon, Min-Jeong;Kim, Seok-Tae
    • The Journal of the Korea institute of electronic communication sciences
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    • v.11 no.9
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    • pp.885-892
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    • 2016
  • The nonlinear sequence generator called the shrinking generator was designed as nonlinear keystream generator composed by two maximum-length LFSRs. The shrunken sequences generated by the shrinking generator are included in the class of interleaved sequences and can be modelled as one of the output sequences of cellular automata (CA). In this paper, we propose a method for synthesizing a 90/150 CA-based sequence generator to generate a family of sequences with the same characteristic polynomial as the shrunken sequences.

Nucleotide Sequences of nodD and nodA from Bradyrhizobium sp. SNU001 (Bradyrhizobium sp. SNU001의 nodD와 nodA의 염기서열)

  • 나영순;심웅섭;안정선
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.189-196
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    • 1993
  • Nucleotide sequences of nodD and nodA from Bradyrhizobium sp. SNUOOI were determined. The open reading frame (ORF) of nodD was 942 bp in length and encoded 314 amino acids. while ORF of nodA, sequence of which is the first one among legume symbionts Bradyrhizobium, was 630 bp and encoded 210 amino acids. The nucleotide sequence of nodD showed 99.4% homology with nodDI of B. japonicum USDAllO. while that of nodA showed 81.5% with B. sp. (Parasponial. At the 5' of nodYAB operon and nodD, consensus nod box sequences composed of 9 bp unit repeated four times and two times respectively were found. Also an A.T-rich sequence was found at 5' of nodD.

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Relationship between Intersequence Pauses, Laying Persistency and Concentration of Prolactin during the Productive Period in White Leghorn Hens

  • Reddy, I.J.;David, C.G.;Singh, Khub
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.686-691
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    • 2005
  • Prolactin is considered to influence the taking of pauses in between ovulatory sequences in White Leghorn hens. Therefore modulating concentrations of prolactin using bromocriptine - a dopamine agonist during early life (17 to 36 weeks of age) could overcome the inhibitory effects of high concentration of prolactin on ovarian activity. The effect of modulation of prolactin concentration on egg production, sequence length and inter sequence pauses were studied by analyzing the oviposition records from 19 to 72 weeks were studied and compared with untreated controls. Bromocriptine administered subcutaneously (100 $\mu$g kg$^{-1}$ body weight or orally through feed (640 $\mu$g day$^{-1}$ bird$^{-1}$) resulted in a steady and sustained decrease in prolactin levels (p<0.01) during and after the withdrawal of treatment up to one reproductive cycle (72 weeks of age). The treated birds had comparatively longer sequences (p<0.01) and fewer pauses (p<0.01). Egg production increased (p<0.01) by fourteen per cent through subcutaneous administration and eleven per cent through oral feeding, over the control birds. It is concluded that the physiological pauses that occur during ovulatory sequences can be disrupted effectively using bromocriptine. Prolactin levels are modulated which may interfere with the follicular recruitment and subsequent oviposition thereby improve egg laying potential of the bird.

Reversible DNA Watermarking Technique Using Histogram Shifting for Bio-Security (바이오 정보보호 위한 히스토그램 쉬프팅 기반 가역성 DNA 워터마킹 기법)

  • Lee, Suk-Hwan;Kwon, Seong-Geun;Lee, Eung-Joo;Kwon, Ki-Ryong
    • Journal of Korea Multimedia Society
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    • v.20 no.2
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    • pp.244-253
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    • 2017
  • Reversible DNA watermarking is capable of continuous DNA storage and forgery prevention, and has the advantage of being able to analyze biological mutation processes by external watermarking by iterative process of concealment and restoration. In this paper, we propose a reversible DNA watermarking method based on histogram multiple shifting of noncoding DNA sequence that can prevent false start codon, maintain original sequence length, maintain high watermark capacity without biologic mutation. The proposed method transforms the non-coding region DNA sequence to the n-th code coefficients and embeds the multiple bits of the n-th code coefficients by the non-recursive histogram multiple shifting method. The multi-bit embedding process prevents the false start codon generation through comparison search between adjacent concealed nucleotide sequences. From the experimental results, it was confirmed that the proposed method has higher watermark capacity of 0.004-0.382 bpn than the conventional method and has higher watermark capacity than the additional data. Also, it was confirmed that false start codon was not generated unlike the conventional method.

Molecular cloning of ribosomal P protein in Toxoplasma gondii and the availability to detect antibody against recombinant protein in toxoplasmosis patients

  • Ahn, Hye-Jin;Kim, Sera;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.41 no.2
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    • pp.89-96
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    • 2003
  • Among the panel of monoclonal antibodies (mAb) against Toxoplasma gondii, mAb of Tg621 (Tg621) clone blotted 38 kDa protein which localized in the cytoplasm of tachyzoites by immunofluorescence microscopy The protein was not released into the parasitophorous vacuole during or after invasion. The cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg621. The full length cDNA sequence was completed with 5’-RACE as 1,592 bp, which contained open reading frame of 942 bp. The deduced amino acid sequence of Tg621 consisted of a polypeptide of 313 amino acids, with significant homology to ribosomal P proteins (RPP) of other organisms especially high to those of apicomplexan species. The expressed and purified TgRPP was assayed in western blot with the sera of toxoplasmosis patients and normal sera, which resulted in the 74.0% of positive reactions in toxoplasmosis patients whereas 8.3% in normal group. Therefore, the antibody formation against TgRPP in toxoplasmosis patients was regarded as specific for T. gondii infection and suggested a potential autoantibody.

Characterization of CaCOP1 Gene in Capsicum annuum Treated with Pathogen Infection and Various Abiotic Stresses

  • Guo, Jia;Seong, Eun-Soo;Wang, Myeong-Hyeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.227-233
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    • 2007
  • We characterized a full-length cDNA of CaCOP1 from pepper. Phylogenetic analysis based on the deduced amino acid sequence of CaCOP1 cDNA revealed high sequence similarity to the COP1 gene in Oryza sativa (84% identity). CaCOP1 shares high sequence identity with regulatory protein in Arabidopsis (84%), constitutively photomorphogenic 1 protein in Pisum sativum (81%) and COP1 homolog in Lycopersicon esculentum (79%). CaCOP1 gene exists single copy in the chili pepper genome. Expression of CaCOP1 was reduced in response to inoculation of non-host pathogens. The expression of this gene under abiotic and oxidative stresses was investigated, including 200 mM NaCl, 200 mM mannitol, cold ($4^{\circ}C$), 100 ${\mu}M$ abscisic acid (ABA), and 10 mM hydrogen peroxide ($H_2O_2$). CaCOP1 was induced significantly 3 h after low temperature treatment but not by dehydration or high salinity. Moreover, CaCOP1 was not induced by plant hormone ABA. These observations suggest that CaCOP1 gene plays a role in abiotic stress and may be belong to ABA-independent regulation system.

Analysis of the Complete Genome Sequence of Zucchini yellow mosaic virus strain A Isolated from Hollyhock

  • Choi, Seung-Kook;Yoon, Ju-Yeon;Sohn, Seong-Han
    • The Plant Pathology Journal
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    • v.23 no.4
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    • pp.245-250
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    • 2007
  • The complete genome sequence of Zucchini yellow mosaic virus stain A (ZYMV-A) isolated from a hollyhock (Althaea rosea) was determined by using RT-PCR with a series of primer sets. The virus genome consisted of 9593 nucleotides (nt), excluding the poly(A) tract at 3' terminus of the virus genome, with 5' and 3' untranslated region of 139 and 211 nt, respectively. The deduced polyprotein of ZYMV-A consisted of 3080 amino acid (aa) residues and was 351 kDa in molecular weight. All proteolytic cleavage sites of the polyprotein of ZYMV-A were compared with those of ZYMV strains, which showed the cleavage sites were conserved among ZYMV strains. The HC-Pro contained the KITC and PTK motifs, and the DAG motif was located at CP ORF of ZYMV-A, suggesting that ZYMV-A is aphid-transmissible. Phylogenetic tree analysis based on the complete genome among ZYMV strains or CP ORFs with other potyviruses showed ZYMV strains formed a distinct group. These results clearly confirmed that ZYMV-A was another distinct strain in ZYMV population at molecular level.

Development and evaluation of semi-nested PCR for detection of the variable lipoprotein haemagglutinin (vlhA) gene of Mycoplasma Synoviae in chicken

  • Pohuang, Tawatchai;Phuektes, Patchara;Junnu, Sucheeva
    • Korean Journal of Veterinary Research
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    • v.60 no.3
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    • pp.109-116
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    • 2020
  • This study aimed to develop a semi-nested polymerase chain reaction assay for the direct detection of Mycoplasma synoviae (M. synoviae) from clinical samples using three newly designed oligonucleotide primers specific to the variable lipoprotein haemagglutinin (vlhA) gene and differentiate M. synoviae field strains based on a nucleotide deletion or the insertion of the proline-rich repeat (PRR) region of the vlhA gene. The developed semi-nested polymerase chain reaction (PCR) assay revealed positive results in 12 out of 100 clinical samples collected from chickens showing lameness and joint swelling. Six positive samples were selected randomly for sequencing, and sequence analysis revealed 96.3-100% nucleotide identities compared to the reference sequences. Phylogenetic analysis showed that sequences of the strains in this study were closely related to WVU1853 (Spain), CK.MS.UDL.PK.2014.2 (Pakistan), and F10-2AS (USA) strains, but they were distinct from the M. synoviae-H vaccine strain sequence. M. synoviae obtained from these samples were identified as types A and C with a length of 38 and 32 amino acids, respectively. These results indicated that the specific and sensitive semi-nested PCR could be a useful diagnostic tool for the direct identification of clinical samples, and the sequence analysis of the partial vlhA gene can be useful for typing M. Synoviae.

Molecular Cloning and Characterization of the Rod Opsin Gene in Olive Flounder Paralichthys olivaceus

  • Kim, Jong-Myoung;Kim, Sung-Wan;Kim, Sung-Koo
    • Fisheries and Aquatic Sciences
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    • v.10 no.1
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    • pp.8-15
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    • 2007
  • Rhodopsin, a dim-light receptor, is a model system for the study of G protein-coupled receptors that transduce extracellular signals into cells. To study the molecular mechanisms of visual systems in fish, the rod opsin gene of olive flounder Paralichthys olivaceus was characterized. The full-length P. olivaceus opsin gene was obtained by PCR amplification of genomic DNA, as well as cDNA synthesis. A comparison of clones obtained from both methods indicated that the olive flounder rod opsin gene lacks introns. Sequence analysis of the opsin gene indicated that it contains a 1,056-bp open reading frame encoding 352 amino acids. The deduced amino acid sequence contains features of typical rod opsins, such as sites for Schiff's base formation (K296) and its counterion (E113), disulfide formation (C110 and C187), and palmitoylation (C322 and C323). An opsin sequence alignment showed the highest similarity between P. olivaceus and Solea solea (95.1%), followed by Hippoglossus hippoglossus (94.5%). An opsin phylogenetic tree revealed a close relationship between olive flounder and teleost rod opsins.