특별강연 : 일본 토양동물학의 연구발전사와 현황

청목순일 ( Jun Ichi Aoki ) 한국토양동물학회 1996 한국토양동물학회지 Vol.1 No.1

Construction of an artificially randomized IgNAR phage display library: screening of variable regions that bind to hen egg white lysozyme.

Ohtani, Maki,Hikima, Jun-ichi,Jung, Tae Sung,Kondo, Hidehiro,Hirono, Ikuo,Aoki, Takashi Springer-Verlag New York Inc 2013 Marine biotechnology Vol.15 No.1

<P>To develop a multi-antigen-specific immunoglobulin new antigen receptor (IgNAR) variable (V) region phage display library, CDR3 in the V region of IgNAR from banded houndshark (Triakis scyllium) was artificially randomized, and clones specific for hen egg white lysozyme (HEL) were obtained by the biopanning method. The nucleotide sequence of CDR3 in the V region was randomly rearranged by PCR. Randomized CDR3-containing segments of the V region were ligated into T7 phage vector to construct a phage display library and resulted in a phage titer of 3.7??10(7) PFU/ml. Forty clones that contained randomized CDR3 inserts were sequenced and shown to have different nucleotide sequences. The HEL-specific clones were screened by biopanning using HEL-coated ELISA plates. After six rounds of screening, nine clones were identified as HEL-specific, eight of which showed a strong affinity to HEL in ELISA compared to a negative control (i.e., empty phage clone). The deduced amino acid sequences of CDR3 from the HEL-specific phage clones fell into four types (I-IV): type I contains a single cysteine residue and type II-IV contain two cysteine residues. These results indicated that the artificially randomized IgNAR library is useful for the rapid isolation of antigen-specific IgNAR V region without immunization of target antigen and showed that it is possible to isolate an antigen-specific IgNAR V region from this library.</P>

Transcriptional regulation of type I interferon gene expression by interferon regulatory factor-3 in Japanese flounder, <i>Paralichthys olivaceus</i>

Ohtani, Maki,Hikima, Jun-ichi,Hwang, Seong Don,Morita, Takahiro,Suzuki, Yoshiaki,Kato, Goshi,Kondo, Hidehiro,Hirono, Ikuo,Jung, Tae-Sung,Aoki, Takashi Elsevier 2012 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol.36 No.4

<P><B>Highlights</B></P><P>► Japanese flounder type I IFN gene was cloned and clustered with Acanthopterygii. ► The poly I:C-responsible region (−634 to −179bp) was found in IFN promoter. ► Transcriptional activity of IFN promoter was enhanced by the flounder IRF3. ► The activity of IFN promoter was induced by RLRs after poly I:C-stimulation.</P> <P><B>Abstract</B></P><P>Type I interferon (IFN) induces the antiviral response in innate immunity. The type I IFN gene cloned from Japanese flounder (<I>Paralichthys olivaceus</I>) has a length of 1189bp and consisting of 5 exons and 4 introns. In a phylogenetic tree of type I IFNs, Japanese flounder grouped with other Acanthopterygii. To gain insight into the transcriptional regulation of IFN gene, the 1.36kb 5′-upstream region including numerous canonical motifs to bind transcription factors [for example, IFN regulatory factor (IRF)] was analyzed. In HINAE cells using a luciferase reporter assay, poly I:C-responsive transcriptional activity was found in the region from −634 to −179bp. This region includes several IRF motifs. In the presence of poly I:C, overexpression of IRF3 and RLR strongly enhanced transcriptional activity. These results suggest that the transcriptional regulation of Japanese flounder type I IFN is regulated by IRF3 after triggering with dsRNA sensors.</P>

Molecular cloning and functional analysis of nucleotide-binding oligomerization domain 1 (NOD1) in olive flounder, <i>Paralichthys olivaceus</i>

Park, Seong Bin,Hikima, Jun-ichi,Suzuki, Yoshiaki,Ohtani, Maki,Nho, Seong Won,Cha, In Seok,Jang, Ho Bin,Kondo, Hidehiro,Hirono, Ikuo,Aoki, Takashi,Jung, Tae Sung Elsevier 2012 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol.36 No.4

<P><B>Highlights</B></P><P>► The gene encoding nucleotide-binding oligomerization domain 1 (NOD1) was cloned in olive flounder. ► NOD1 was expressed in all fish tissues examined. ► NOD1 mRNA levels was elevated in fish infected with <I>Edwardsiella tarda</I>, <I>Streptococcus iniae</I>, or VHSV. ► The inhibition of <I>E. tarda</I> and the increase of IL-1β levels were observed in NOD1 over-expressing cells.</P> <P><B>Abstract</B></P><P>The gene encoding nucleotide-binding oligomerization domain 1 (NOD1) was cloned from olive flounder (<I>Paralichthys olivaceus</I>) and the role played by NOD1 during <I>Edwardsiella tarda</I> infection was evaluated. The complete open reading frame of NOD1 was 2820bp in length, encoding a 939-amino acid polypeptide. The NOD1 protein contains three conserved domain structures including C-terminal LRRs, a central NACHT motif, and an N-terminal CARD domain, which show similarities of 49–74% to those of other vertebrate counterpart proteins. NOD1 expression was observed in all fish tissues examined, and the levels increased in olive flounder infected with <I>E. tarda</I>, <I>Streptococcus iniae</I>, or viral hemorrhagic septicemia virus (VHSV). When hirame natural embryo (HINAE) cells over-expressing NOD1 were infected with <I>E. tarda</I>, bacterial growth was inhibited, and the IL-1β transcript level increased compared to that of the control. These findings imply that NOD1 plays an important role in response to <I>E. tarda</I> infection of olive flounder.</P>

The Oribatid Mites (Acari: Cryptostigmata) of Korea (4) On the femoralis-group of the genus Carabodes C.L, Koch, 1836

Park, Seong-Sik,Aoki, Jun-ichi 한국곤충학회 1986 Korean journal of entomology Vol.16 No.1

한국산 날개응애류중에서 검둥이응애과(Caratodiidae)에 속하는 무늬검둥이응애 (Carabodes transversarius sp. n.)와 어깨점둥이응애 (C. tsushimaensis dorsalis subsp. n.)를 신종으로 기재하고 대마도 검둥이응애 (C. tsushimaensis Aoki, 1970)를 미기록종으로 기록하며 Caradodes속의“femoralis group”의 종검색표를 작성하였다. Two new species and one unrecorded species of Korean oribatid mites belonging to the genus Car-aboder C. L. Koch, 1836 (Carabodiidae) are described. They are C. transversarius sp. nov., C. tsu shimaensis dorsalis subsp. nov. and C. tsushimaensis Aoki, 1970. A Key to the species of“femoralis group”of the genus Caradodes is presented.

Three New Species of Oribatid Mites (Acarina, Oribatei) from Korea

Choi, Seong-Sik,Aoki, Jun-ichi 한국곤충학회 1993 Entomological Research Vol.23 No.1

본 연구는 한국산 날개응애 3신종, 동양판지게응애 (Oribatella orientalis) 문산곰보응애 (Xenillus moonsani) 한국도포응애 (Perxylobates coreanus)를 기재하였다. 동양판지게응애는 등판, 익상돌기 및 복판의 표면이 거의 규칙적인 그물무늬로 덮여있고, 머리끝의 양쪽에 삼지창모양의 부속기가 붙어있는 것이 특징이다. 문산곰보응애는 길고 넓은 지게의 모양이 특징 적이며 지게털이 지게의 복면쪽에 나 있다. 한국도포소매응애는 가슴판털에 가시털이 분지되어 있고, 지금까지 보고된 민목도포응애속(신칭 : Perxylobates)의 다른종들은 생식판털이 5쌍인데 반하여 이 종은 4쌍인 것이 특징이다. Three new species of oribatid mites, Oribatella orientalis sp. nov., Xenillus moonsani sp. nov., and Perxylobates coreanus sp. nov., are described from Korea. In O. orientalis, the surface structures of notogaster, pteromorphae, and ano-genital plates are distinctly sculptured with almost regular pattern of polygonal network and the rostrum has on either side a blade-like appendage with tricuspid tip. The epimeral setae, 3b, c and 4a, b are distinctly long and thick, of which 4b is even more distinct. X. moonsani is characteristic in long and wide lamellae. The lamellar setae are inserted on ventral side of lamellae. P coreanus differs from the known species in the barbed epimeral setae and four pairs of genital setae instead of five in other members of the genus Perxylobates.

韓國產 날개응애類(3) Defectamerus 屬의 新種에 關하여

SEONG SIK CHOI,JUN-ICHI AOKI 한국응용곤충학회 1985 한국응용곤충학회지 Vol.24 No.3

Defectamerus soonkii sp. nov. and D. crassisetiger coreanus subsp. nov. belonging to the family Ameridae are described. Key to the species of the genus Defectamerus Aoki is presented. 한국산(韓國産) Defectamerus속(屬)의 1 신종(新種), 1 신아종(新亞種) 및 종검색표(種檢索表)를 작성(作成)하였다. 신종(新種)은 순기민동정응애(D. soonkii sp. nov.)와 신아종(新亞種)으로 한국민동정응애(D. crassisetiger coreanus subsp. nov.)를 기재(記載)하였다.

Molecular cloning and antiviral activity of IFN-β promoter stimulator-1 (IPS-1) gene in Japanese flounder, <i>Paralichthys olivaceus</i>

Simora, Rhoda Mae C.,Ohtani, Maki,Hikima, Jun-ichi,Kondo, Hidehiro,Hirono, Ikuo,Jung, Tae Sung,Aoki, Takashi Elsevier 2010 FISH AND SHELLFISH IMMUNOLOGY Vol.29 No.6

<P><B>Abstract</B></P><P>The mitochondrial adaptor, IFN-β promoter stimulator-1 (IPS-1), also known as MAVS/VISA/Cardif, plays a key role in the signal transduction of the RIG-1/MDA5 pathway to induce the production of interferons (IFNs) and other cytokines. In the present study, Japanese flounder (<I>Paralichthys olivaceus</I>) IPS-1 cDNA was cloned from Japanese flounder spleen using PCR-based methods. The full-length cDNA has 2235 nucleotides and encodes a polypeptide of 641 amino acids. The putative Japanese flounder IPS-1 protein contains an N-terminal CARD-like domain, a central proline-rich domain, a C-terminal transmembrane domain, and exhibits similarity to other teleost counterparts ranging from 20% to 34%. Semi-quantitative RT-PCR showed that Japanese flounder IPS-1 mRNA was expressed in all tissues examined. The expression level of flounder IPS-1 gene was unchanged in viral hemorrhagic septicemia virus (VHSV)-infected kidney as measured by quantitative real-time PCR (Q-PCR). In addition, Japanese flounder IPS-1-overexpressing cells were protected against hirame rhabdovirus (HIRRV) and VHSV infection as manifested by the delayed appearance of cytopathic effect (CPE) and decreased viral titers. Expression of IFN-inducible genes including Mx, ISG15 and IRF3 were also induced in the IPS-1-overexpressing cells. These results suggest that Japanese flounder IPS-1 is involved in the antiviral immunity as a one of the adaptors in fish IFN-activation pathway.</P>

Comparative Sequence Analysis of a Multidrug-Resistant Plasmid from Aeromonas hydrophila

del Castillo, Carmelo S.,Hikima, Jun-ichi,Jang, Ho-Bin,Nho, Seong-Won,Jung, Tae-Sung,Wongtavatchai, Janenuj,Kondo, Hidehiro,Hirono, Ikuo,Takeyama, Haruko,Aoki, Takashi American Society for Microbiology 2013 Antimicrobial agents and chemotherapy Vol.57 No.1

<B>ABSTRACT</B><P>Aeromonas hydrophilais a pathogenic bacterium that has been implicated in fish, animal, and human disease. Recently, a multidrug resistance (MDR) plasmid, pR148, was isolated fromA. hydrophilaobtained from a tilapia (Oreochromis niloticus) farm in Thailand. pR148 is a 165,906-bp circular plasmid containing 147 coding regions showing highest similarity to pNDM-1_Dok1, an MDR plasmid isolated from a human pathogen. pR148 was also very similar to other IncA/C plasmids isolated from humans, animals, food, and fish. pR148 contains a mercuric resistance operon and encodes the complete set of genes for the type 4 secretion system. pR148 encodes a Tn<I>21</I>type transposon. This transposon contains the drug resistance genes<I>qacH</I>,<I>bla</I>OXA-10,<I>aadA1</I>, and<I>sul1</I>in a class 1 integron;<I>tetA</I>and<I>tetR</I>in transposon Tn<I>1721</I>; and<I>catA2</I>and a duplicate<I>sul1</I>in a locus showing 100% similarity to IncU plasmids isolated from fish. The<I>bla</I>OXA-10and<I>aadA1</I>genes showed 100% similarity to those from theAcinetobacter baumanniiAYE genome. The similarity of pR148 to a human pathogen-derived plasmid indicates that the plasmids were either transferred between different genera or that they are derived from a common origin. Previous studies have shown that IncA/C plasmids retain a conserved backbone, while the accessory region points to lateral gene transfer. These observations point out the dangers of indiscriminate use of antibiotics in humans and in animals and the necessity of understanding how drug resistance determinants are disseminated and transferred.</P>

Molecular characterization, expression and functional analysis of a nuclear oligomerization domain proteins subfamily C (NLRC) in Japanese flounder (<i>Paralichthys olivaceus</i>)

Unajak, Sasimanas,Santos, Mudjekeewis D.,Hikima, Jun-ichi,Jung, Tae-Sung,Kondo, Hidehiro,Hirono, Ikuo,Aoki, Takashi Elsevier 2011 Fish & shellfish immunology Vol.31 No.2

<P><B>Abstract</B></P><P>Pattern recognition receptors (PRRs) are involved in the effective innate defense against several microbes. Here, we identified a nucleotide-oligomerization domain (NOD)-like receptor subfamily C (NLRC) from Japanese flounder (<I>Paralichthys olivaceus</I>). Full-length transcript of JfNLRC is composed of 3976bp encoding a protein of 1175 deduced amino acid residues. The presence of a signature nucleotide-binding domain (NACHT) and leucine-rich repeated domain (LRR) suggested that the protein is a member of the NLR family. Interestingly, its C-terminus presents an extra PRY/SPRY (B30.2) domain similar to fish in the Trim (finTrim) family. A phylogenic tree of JfNLRC revealed that full-length JfNLRC diverged from the NOD1 and NOD2 clusters, and the NACHT domain in JfNLRC was clustered within the NLRC3 group. Stimulation by formalin-killed <I>Edwardsiella tarda</I>, <I>Streptococcus iniae</I>, and lipopolysaccharide (LPS) showed that the JfNLRC expression was raised a few hours after stimulation, suggesting this novel protein is involved in the immediate response against both Gram-positive and Gram-negative bacteria. Furthermore, the IL-1β mRNA expression level in JfNLRC-over-expressing HINAE cells was significantly increased, when compared to a control, after LPS-stimulation and <I>E. tarda</I> infection. These results suggested that JfNLRC probably induced IL-1β gene expression mediated by LPS-stimulation.</P> <P><B>Highlights</B></P><P>► NLRC mRNA in Japanese flounder was cloned. ► The phylogenetic tree was analyzed and evolutionary conservation was clarified. ► The tissue distribution of NLRC was measured. ► NLRC transcripts were increased by stimulation with formalin-killed bacteria and LPS. ► NLRC probably mediates induction of IL-1b mRNA expression through LPS-stimulation.</P>