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First Record of Adults of Prosimulium and Twinnia (Diptera: Simuliidae) from Korea

  • Kim, Sam-Kyu
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.4
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    • pp.285-295
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    • 2020
  • Adults of Prosimulium kiotoense and Twinnia japonensis were collected and reported from Korea for the first time. Since adult black flies are notorious for morphological homogeneity particularly in female, larval and pupal characters have been mainly used to identify them. Adults of P. kiotoense can be identified by the following combination of characteristics: Adult, wing with radial sector (Rs) branched into R2+3 and R4+5; hind leg basitarsus without calcipala; first tarsomere without pedisulcus. Female, claw without basal thumblike lobe; hypogenial valve elongate, convex, heavily sclerotized medially, posterior end touching each other, space between valves rhomboid. Male, claw with basal thumblike lobe; ventral plate keel shaped; gonostylus with 2-3 spinules. Adults and pupa of T. japonensis can be identified by the following combination of characteristics: Adult, antenna with 7 flagellomeres. Female, hypogenial valve broad, posterior end of valve not touching each other; cercus elongate, subquadrate; spermatheca slightly wider and long, round. Male, claw with basal thumblike lobe; gonostylus with 1 spinule; ventral plate flat. Pupa, gill of 16 filaments, arising from 3 swollen stalks; abdominal tergites without spine combs except tergites III and IV with small recurved hooks; terminal spine well developed, wavy shaped.

Expression of Fusion Protein with Autographa californica Nuclear Polyhedrosis Virus Polyhedrin and Bacillus thuringiensis cryIA(c) Crystal Protein in Insect Cells (곤충세포주에서 Autographa californica 핵다각체병 바이러스의 다각체 단백질과 Bacillus thuringiensis cryIA(c) 내독소 단백질의 융합 단백질 발현)

  • 제연호;진병래;박현우;노종열;장진희;우수동;강석권
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.341-350
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    • 1997
  • We have now constructed a novel recombinant baculovirus producing fusion protein with Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin and Bacillus thuringiensis(Bt) cryIA(c) crystal protein. The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The N-terminal of cryIA(c) gene of Bt subsp. kurstaki HD-73 was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front of intact polyhedrin gene or by insertion into the HindIII site in polyhedrin gene. The recombinant baculoviruses were named as BtrusI or BtrusII, respectively. Although single transcript from the fusion protein gene was apparently observed. BtrusI was produced the two proteins, 92 kDa fusion protein and only polyhedrin. In addition, fusion protein produced by BtrusI did not form polyhedra. Interestingly, however, the cells infected with BtrusII did not show a 33 kDa polyhedrin band as a cells infected with BtrusI. Cells infected with BtrusII were only produced fusion protein, but the polyhedra formed by fusion protein was not observed. To determine the insecticidal toxicity of fusion protein, therefore, Sf9 cells infected with BtrusI were inoculated to Bombyx mori larvae. Sf9 cells infected with BtrusI that expressed the fusion protein caused larval mortality although the insecticidal toxicity was low. In conclusion, our results clearly demonstrated that the fusion protein with polyhedrin and Bt cryIA(c) crystal protein have a insecticida toxicity.

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Effect of RFLP Marker of the Mitochondrial DNA D-Loop Region on Milk Production in Korean Cattle (한우 Mitochondrial DNA D-Loop 영역의 RFLP Marker가 산유량에 미치는 영향)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
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    • v.25 no.2
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    • pp.218-225
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    • 2005
  • This study was performed to determine sequence variation and RFLP of the mt DNA D-loop region using Southern blot hybridization analysis and to develop mt DNA marker affecting milk production traits in Hanwoo cows. The PCR was used to amplify an 1142 bp fragment within the D-loop region of mt DNA using specific primers. Mt DNA were digested with seven restriction enzymes and hybridized using DIG-labeled D-loop probe. The mt DNA RFLP polymorphisms were observed in the four enzymes, BamHI, RsaI, XbaI and HpaII. Nucleotide substitutions were detected at positions 441 (G/C), 469 (T/C), 503 (C/T), 569 (G/A), 614 (C/A) and 644 (C/T) of the mt DNA D-loop region between two selected lines. Significant relationship between the XbaI RFLP type and breeding value was found(p<0.05). Cows with A type had higher estimated breeding values than those with B type (P<0.05) between high and low milk production lines. Therefore, the RFLP marker of mt DNA could be used as a selection assisted tool for individuals with high milk producing ability in Hanwoo.

Effects of Achyranthes Radix and Electroacupuncture on Type II Collagen-Induced Arthritis (류마티스 관절염에서 우슬과 전침자극의 항관절염 효과)

  • Kim, Gye-Yeop;Seong, Rak-Seon;Kim, Young-Eok;Chang, Mee-Kyung;Yu, Young-Dae;Choi, Ki-Bok;Jeong, Hyun-Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.2
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    • pp.425-431
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    • 2007
  • The purpose of this study was to observe the effects of Achyranthes Radix(AR) and electroacupuncture(EA) in rats with rheumatoid arthritis induced by type II collagen for 28 days. Control group was daily administered 0.9% NaCl 0.5 $m{\ell}$, Group I was daily administered 0.9% NaCl 0.5 $m{\ell}$ to arthritic rats, Group II was orally administered with Achyranthes Radix 500 mg/kg 0.5 $m{\ell}$ to arthritic rats. Group III was given 2 Hz EA of chok samni acupoint(ST36) in the test group for 30 min/days to arthritic rats. Group IV was daily orally administered with Achyranthes Radix 500 mg/kg 0.5 $m{\ell}$ and 2 Hz EA of chok samni acupoint(ST36) in the test group for 30 min/days to arthritic rats. This studies have been designed to evaluate the hind paw edema, assessment of arthritis indices, analgetic effects by analysis of blood chemistry(WBC, CRP, ALP, AST). In each group, histologic observations, Safranin O-fast green stain were observed and analyzed. The following results were obtained. Group II, III, IV were significantly decreased arthritis indices and the rate of paw edema compared with Group I . Especially group IV was the most significantly decreased. The WBC, CRP, AST, ALT was that Group II, III, IV were significantly decreased compared with Group I . In conclusion, Achyranthes Radix and Ea contribute to the improvement of blood chemistry and change in safranin O-fast green by knee joint of arthritic rats.

Development of Restriction Fragment Length Polymorphism(RELP) Markers in Silkworm, Bombyx mori (누에 RFLP(제한단편 다형현상)마커 개발)

  • 고승주;김태산;이영승;황재삼;이상몽
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.96-104
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    • 1997
  • A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

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Cloning of the Polyhedrin Gene-Containing DNA Fragment of Hyphantria cunea Nuclear Polyhedrosis Virus (흰불나방 핵다각체바이러스 다각체단백질 유전자포함 절편의 클로닝)

  • 박호영;진병래;박순식;김정일;깅석권
    • Korean journal of applied entomology
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    • v.32 no.1
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    • pp.51-60
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    • 1993
  • The polyhedrin gene-containing DNA fragment of Hyphantria cunea nuclear polyhedrosis virus (HcNPV) was localized by southern hybridization with Autographa california CPA EcoRI-I fragment (7.3 kb), Bombyx mori NPV PatI-F fragment (7 kb) and synthetic oligonucleotide(30-mer) as probes. the PstI-L(5.3 kb) fragment of HcNPV was cloned to E. coli and the plasmid of the fragment was named as pHcP-L(8.0 kb). The pHcP-L was physically mapped and subcloned to E. coli as pHcP-L1(4.7 kb), pHcP-L2(7.1 kb), pHcP-L3(5.3 kb), pHcP-L4(4.2 kb) and pHcP-L5(4.5 kb).

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Study on development of DNA probe for identification of Prevotella intermedia G8-9-3 (Prevotella intermedia G8-9K-3을 동정할 수 있는 DNA 프로브의 개발에 관한 연구)

  • Bak, Jong-Sung;Kim, Se-Hoon;kim, Dong-Ki;Seong, Jin-Hyo;Kim, Byung-Ock;Kim, Jung-Ki
    • Journal of Periodontal and Implant Science
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    • v.32 no.2
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    • pp.281-290
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    • 2002
  • The purpose of this study is to develop species-specific DNA probe for detection and identification of Prevotella intermedia (P. intermedia) G8-9K-3. This study procedure includes (1) whole-genomic DNA extraction of P. intermedia G8-9K-3 (2) construction of the genomic DNA library, (3) screening of strain-specific DNA probe by reverse dot hybridization, (4) confirmation of strain-specific DNA probe by Southern blot hybridization, (5) determination of nucleotide sequences of strain-specific DNA probe. Twenty-eight recombinant plasmids containing Hind III-digested DNA fragments of P. intermedia G8-9K-3 were obtained. Reverse Dot Hybridization and Southern blot analysis data showed that one of them, Pig3, could be P. intermedia G8-9K-3-specific DNA probe. This datum indicates that this Pig3 DNA probe could be useful in detection and identification of the P. intermedia G8-9K-3 strain.

Virulence-associated plasmids of Salmonella spp. isolated from animals in Korea (동물에서 분리된 Salmonella균의 병원성 관련 Plasmid에 관한 연구)

  • Choi, Won-pil;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.32 no.3
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    • pp.369-376
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    • 1992
  • This paper dealt with plasmid DNA profile in 98 Salmonella(S) isolated from pigs and cattle sources in Taegu, Gyeongbook and Gyeongnam during the period from 1984 to 1987. Also we were studied for restriction enzyme analysis of the plasmid DNA, and mouse infection, Sereny test and normal setum resistance test in guinea pig for S typhimurium and S enteritidis harbored or cured 60 megadalton(Md) plasmid and 36 Md plasmid, respectively. Of the 13 Salmonella isolated from cattle, 7 Salmonella harbored one or more plasmids and molecular sizes of the large plasmids were 60 Md for S typhimurium and 36 Md for S enteritidis. Of the 85 Salmonella isolated from pigs, 47 Salmonella were confirmed as being one or more plasmids, and all the S typimurium stains harbored 60 Md plasmid. In enzyme digestion with 8 types of restriction endonuclease for 60 Md plasmid DNA of S typhimurium, cleavage patterns were varied to enzymes, and the DNA was segmented into 4 to 15 fragments. In restriction enzyme analysis of 36 Md plasmid DNA obtained from four strains of S. enteritidis, the DNA showed the same cleavage patterns obtained with Eco RI, Hind III and Bam H I, and was segmented into 3 to 5 fragments. In virulence for mice by measuring the 50% lethal dose ($LD_{50}$), the $LD_{50}$ values obtained for 60 Md virulence-associated plasmid harbored strains of S typhimurium and 36 Md virulence-associated plasmid of S enteritidis were up to $10^4$-fold lower than the values obtained for the plasmid-cured strains of the same serotype. Only the plasmid harbored strains were resistant to the bactericidal activity of 90% guinea pig serum, and only they gave positive responses in sereny test. We suggested that their plasmid DNA might be associated with virulence for mice.

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Polymorphism of mitochondrial DNA in Jindo dogs and Japanese mongrels dogs (DNA 다형(多型)에 있어서 진도견(珍島犬)과 잡종견(雜種犬)과의 비교(比較))

  • Han, Bang-keun;Kim, Joo-heon;Kang, Ju-won;Ikemoto, Shigenori
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.43-51
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    • 1993
  • Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

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White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.7-13
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    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.