• Title/Summary/Keyword: resistant gene

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Identification of potential molecular markers for disease resistance in giant gourami through major histocompatibility complex (MHC) II gene analysis

  • Ikhsan Khasani;Rita Febrianti;Sularto;Wahyu Pamungkas;Keukeu Kaniawati Rosada
    • Fisheries and Aquatic Sciences
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    • v.27 no.3
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    • pp.159-170
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    • 2024
  • Research to obtain molecular markers related to the major histocompatibility complex (MHC) gene in both strains of gourami is essential to increase the success of the selection program of disease resistance traits. Using a completely randomized design (CRD), the challenge test consists of four treatments and seven replications. The treatment was Jambi gourami injected with PBS (KJ), Kalimantan gourami injected with PBS (KK), Jambi strain injected with Aeromonas hydrophila (GJ), and Kalimantan strain injected with A. hydrophila (GK). The GJ population was more resistant to A. hydrophila than the GK population. The MHC II gene was detected in both test strains (GJ and GK), both resistant and susceptible fish. However, there were differences in the results of amplifying the MHC II gene in susceptible and resistant fish. Two DNA fragments approximately 400 and 585 bp were detected in the genome of susceptible fish, while in the genome of susceptible fish, only one DNA fragment was detected (400 bp). Therefore, the MHC II gene fragment with a size of about 585 bp can be used as a potential candidate for specific molecular markers to obtain resistance to A. hydrophila bacteria in the giant gourami.

Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis (세균성 피부염 개에서 분리된 Staphylococcus pseudintermedius에서 항생제 감수성 검사와 내성 유전자 획득의 비교)

  • Jang, Hye-Jin;Son, Hyoung-Won;Kang, Hyo-Min;Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.158-161
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    • 2015
  • Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.

Antibiotic-Resistance Profiles and the Identification of the Ampicillin-Resistance Gene of Vibrio parahaemolyticus Isolated from Seawater (해수에서 분리한 장염비브리오의 항생제 내성 및 암피실린 내성 유전자의 동정)

  • Lee, Kuen-Woo;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.6
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    • pp.637-641
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    • 2010
  • The antibiotics-resistance profiles of 28 strains of Vibrio parahaemolyticus isolated from seawater were investigated. All of the strains studied were resistant to ampicillin (100%), but susceptible to 12 other antibiotics. The minimum inhibitory concentration (MIC) of V. parahaemolyticus to ampicillin was as high as $1,024-2,048\;{\mu}g{\cdot}mL^{-1}$. The phenotype of strain 8 changed from ampicillin-resistant to susceptible with an in-frame deletion mutant of VPA0477, a putative ${\beta}$-lactamase gene, and the MIC for ampicillin of the mutant strain was $1{\mu}g{\cdot}mL^{-1}$. In conclusion, our findings suggest that the VPA0477 gene acts as a ${\beta}$-lactamase in ampicillin-resistant V. parahaemolyticus strains.

Differentially Expressed Genes Related to Cold-resistance in Barley (Hordeum vulgare L. cv. Nagaoka)

  • Chun, Jong Un;Park, Jeong-Seon;Bae, Chang-Hyu;Shin, Jeong-Sheop
    • Korean Journal of Breeding Science
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    • v.41 no.2
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    • pp.73-83
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    • 2009
  • To investigate genes related to vernalization and cold- resistance in barley (Hordeum vulgare L. cv. Nagaoka), differentially expressed genes were identified from cold-resistant barley leaves with suppression subtractive hybridization (SSH) and Northern blot analyses. The nucleotide and the deduced amino acid sequences of the putative gene products were compared. The bvrn-7 showed high homology(84%) with gene related to vernalization, and the bvrn-3, bvrn-12, bvrn-28, bvrn-29 and bvrn-36 related to cold-resistant genes had high identity of 88~98% with low temperature-induced genes. The results indicate that the 6 genes were closely related to vernalization and cold-resistance during low temperature treatment.

Studies on the Factors Influencing the Transformation in Escherichia with pBR322 DNA (Escherichia coli의 pBR322 DNA 형질전환에 관여하는 인자에 관한 연구)

  • Yoo, Han-sang;Mah, Jum-sool
    • Korean Journal of Veterinary Research
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    • v.24 no.1
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    • pp.40-49
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    • 1984
  • To investigate the factors influencing the artifical transformation in Escherichia coli, E. coli C600 was transformed by pBR322 DNA with tetracycline and ampicillin resistant gene purified by CsCl-Etbr equilibrium density gradient centrifugation from E.coli HB 101. The influencing factors in the transformation such as concentration of calcium chloride, time of ice incubation, temperature and time of heat shock, time of gene expression, effects of plasmid DNA concentration and adding time were examined in these experiments. The results obtained were as follows; 1. The highest transformation frequency was observed in the treatments of 100 mM $CaCl_2$ before heat shock and the treatment of $CaCl_2$ was essential step in the process of E. coli transformation. 2. The highest transformation frequency was observed in the treatment of heat shock at $42^{\circ}C$ for 4 min. or $37^{\circ}C$ for 6 min., but the prolonged heat shock resulted a decreased transformation frequency. 3. Treatments of ice incubation at $0^{\circ}C$ for 45 min. before heat stocks or at $0^{\circ}C$ for 30min. after heat shock resulted an increased transformation frequency. 4. There was a linear relationship between DNA concentration and transformation frequency at the concentration of $8{\times}10^3$ recipient cells. The highest transformation frequency reached in carte of 7 mcg of donor DNA, but above 1 mcg of DNA concentration, transformation frequency was not remarkably increased. Addition of donor DNA just after the treatment of $CaCl_2$ was the best. 5. The best condition of gene expression at $37^{\circ}C$ were 40min. for TC-resistant gene and 100min. for AP-resistant gene. TC-resistant gene was higher in the transformation frequency and faster in the gene expression time than AP-resistant gene. In these results, the best conditions for the transformation of E. coli C 600 with pBR322 DNA were: treatment with 100mM $CaCl_2$, ice incubation at $0^{\circ}C$ for 45 min, heat shock at $42^{\circ}C$ for 4 min., 30 min. of ice incubation and incubation at $37^{\circ}C$ for 100min. for gene expression in that order.

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A Rapid Detection of Methicillin-Resistant Staphylococci by Polymerase Chain Reaction (Polymerase Chain Reaction을 이용한 Methicillin-resistant Staphylococci의 신속 검출)

  • 박진숙;박영진
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.306-311
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    • 2002
  • PCR of the mecA gene for the rapid detection of methicillin-resistant staphylococci was perfomed and compared with the antibiotic sensitivity test. A total of 43 strains of staphylococi from clinical specimens were used in this study. An antibiotic sensitivity test by the agar dilution method of NCCLS (The National Commitee for Clinical Laboratory Standard) was performed for the strains. Among them, 39 isolates were methicillin-resistant (MRS), and 4 isolates were methicillin-susceptible (MSS). With the exception for one strain (Staphylococcus cohnii, HRC2-4), all MRS strains amplified the expected 533 bp fragments of the mecA gene by PCR, However, one strain (Staphylococcus aureus, HSA1-10) that was classified as a sensitive strain by the antibiotic sensitivity test was mecA positive by PCR. All 35 methicillin-resistant Staphylococcus aureus (MRSA) strains were mecA positive, but overall, concordance between the results of the mecA PCR and antibiotic sensitivity test was 95.6%.

Screening of Bacterial Leaf Blight Resistance Genes (xa5, xa13, Xa21) using Sequence Tagged Site (STS) Marker in Korean Varieties and Landraces

  • Kim, Young-Chang;Park, Yong-Jin;Ma, Kyung-Ho;Lee, Jung-Ro;Kim, Chang-Young;Choi, Jae-Eul;Kang, Hee-Kyoung
    • Plant Resources
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    • v.7 no.3
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    • pp.187-194
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    • 2004
  • Sequence-tagged site (STS) markers tightly linked to the bacterial leaf blight (BLB) resistance genes, xa5, xa13 and Xa21, were used in this study. A survey was conducted to find polymorphisms between the resistant and susceptible germplasm in rice. 500 of Korean varieties and 100 of landraces were evaluated in this study. STS marker, RG207 was used to having xa5 resistance gene of rice germplasm. 27 varieties of Korean germplasm showed resistant for xa5 gene. The RG136 an xa-13 marker resulted in a single band of approximately 1kb in all the rice accessions studied. In order to detect polymorphism, digestion of the polymerase chain reaction (PCR) product was performed using a restriction enzyme Hinf Ⅰ. The resistant lines resulted in two bands 0.5kb on digestion with Hinf Ⅰ, while the same enzyme did not digest the PCR product of susceptible lines. No polymorphism was detected in Korean varieties and landraces, indicating that they probably do not contain xa13 gene. pTA248 an Xa-21 marker detected a band of 1kb in the resistant lines and bands of either 750bp or 700bp in the susceptible lines. Among germplasm tested, there are no varieties and landraces with Xa21 resistant gene. The results of the germplasm survey will be useful for the selection of parents in breeding programs aimed at transferring these bacterial blight resistance genes from one varietal background to another.

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HPAI-resistant Ri chickens exhibit elevated antiviral immune-related gene expression

  • Thi Hao Vu;Jubi Heo;Yeojin Hong;Suyeon Kang;Ha Thi Thanh Tran;Hoang Vu Dang;Anh Duc Truong;Yeong Ho Hong
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.13.1-13.11
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    • 2023
  • Background: Highly pathogenic avian influenza viruses (HPAIVs) is an extremely contagious and high mortality rates in chickens resulting in substantial economic impact on the poultry sector. Therefore, it is necessary to elucidate the pathogenic mechanism of HPAIV for infection control. Objective: Gene set enrichment analysis (GSEA) can effectively avoid the limitations of subjective screening for differential gene expression. Therefore, we performed GSEA to compare HPAI-infected resistant and susceptible Ri chicken lines. Methods: The Ri chickens Mx(A)/BF2(B21) were chosen as resistant, and the chickens Mx(G)/BF2(B13) were selected as susceptible by genotyping the Mx and BF2 genes. The tracheal tissues of HPAIV H5N1 infected chickens were collected for RNA sequencing followed by GSEA analysis to define gene subsets to elucidate the sequencing results. Results: We identified four differentially expressed pathways, which were immune-related pathways with a total of 78 genes. The expression levels of cytokines (IL-1β, IL-6, IL-12), chemokines (CCL4 and CCL5), type interferons and their receptors (IFN-β, IFNAR1, IFNAR2, and IFNGR1), Jak-STAT signaling pathway genes (STAT1, STAT2, and JAK1), MHC class I and II and their co-stimulatory molecules (CD80, CD86, CD40, DMB2, BLB2, and B2M), and interferon stimulated genes (EIF2AK2 and EIF2AK1) in resistant chickens were higher than those in susceptible chickens. Conclusions: Resistant Ri chickens exhibit a stronger antiviral response to HPAIV H5N1 compared with susceptible chickens. Our findings provide insights into the immune responses of genetically disparate chickens against HPAIV.

Characteristics of Cadmium-Resistant Bacillus subtilis DT134 (Bacillus subtilis DT134의 카드뮴 저항성)

  • 윤경표
    • KSBB Journal
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    • v.13 no.4
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    • pp.383-390
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    • 1998
  • Bacillus subtilis DT134 was resistant to 50-fold higher concentration of cadmium ions (Cd2+) than cadmium-sensitive B. subtilis BD224 in Luria Broth (LB) medium. Minimal inhibition concentration test in LB agar plates also showed similar results. The elevated cadmium resistance of B. subtilis DT134 strongly suggested a possible existence of cadmium resistance gene in it. Southern blot with Staphylococcus aureus cadA gene fragment (757 bp NlaIV-XmnI cadA DNA fragment) as probe was carried out to test the existence and similarity of the gene. In high stringency condition, there was no detectable signal, but in low stringency, a strong signal specific to the cadA probe could be detected. These results strongly suggested that there was some similarity between total DNA of B. subtilis DT134 and S. aureus pl258 in terms of cadmium resistance gene and the resistance mechanism might be an efflux mechanism. The subsequent efflux experiment showed that the cadmium resistance mechanism of B. subtilis DT134 was also due to the efflux of cadmium.

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Effect of ozone treatment on the expression of a foreign gene in transgenic poplar subclones (형질전환 포플라 subclone의 도입유전자 발현에 대한 오존처리의 영향)

  • 설일환;신동일
    • Journal of Life Science
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    • v.7 no.3
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    • pp.172-175
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    • 1997
  • Transgenic hybrid poplar subclones containing herbicide glyphosate resistant gene (aroA) were treated with ozone at the concentration of 100 nL L$^{-1}$ for 6 hr for 5 consecutive days. The foreign gene expression in leaves of all treated plants was reduced both at transcriptional and translational levels confirmed by Northern and Western blot analysis, respectively, as compared to non-treated control plants. These results indicated that the expression of foreign gene in transgenic plants could be affected by the environmental stresses. Thus, the performance of transgenic plants cultivated on field conditions may be lower than they are expected.

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