• Title/Summary/Keyword: InvA Gene

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Investigation on antimicrobial resistance genes of Salmonella spp. isolated from pigs and cattle (소와 돼지유래 살모넬라속균의 약계내성유전자의 특성에 관한 연구)

  • Lee, Woo-Won;Jung, Byeong-Yeal;Lee, Gang-Rok;Lee, Dong-Soo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.32 no.3
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    • pp.227-239
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    • 2009
  • At the present study, it was aimed to detect virulence genes and antimicrobial resistance genes among 102 strains of 12 Salmonella serotypes isolated from pigs and cattle. In polymerase chain reaction (PCR), invA was detected from all strains of Salmonella spp., spvC was detected from Salmonella enterica serotype Enteritidis (S. Enteritidis) (100%), S. Bradenburg (75%), and S. Typhimurium (20.4%). Drug resistance related genes of 12 types were detected from all strains. TEM ($bla_{TEM}$) gene was detected from 51 (92.7%) of 55 $\beta$-lactams (54 ampicillin or 1 amoxicillin) resistance strains. 55 (100%) of 55 chloramphenicol resistance strains, 3 (100%) of 3 gentamicin resistance strains and 5 (100%) of 5 kanamycin resistance strains did contain cml, aadB, and aphA1-Iab, respectively. strB (89.9%), strA (88.4%), aadA2 (84.1%) and aadA1 (72.5%) were detected from 69 streptomycin resistance strains. sulII and dhfrXII were detected from 49 (100%) of 49 sulfamethoxazole/trimethoprim resistance strains, but sulI was not detected. tetA (97.9%) and tetB (21.6%) were detected from 97 tetracycline resistance strains. int gene was detected from 58 (56.9%) of 102 strains. 54 S. Typhimurium of 102 Salmonella spp. were attempted to detect drug resistance genes. TEM was detected from 44 (95.7%) of 46 $\beta$-lactams (45 ampicillin or 1 amoxicillin) resistance strains. cmlA was detected from 51 (100%) of 51 chloramphenicol resistance strains. aadA2 (100%), strA (100%), strB (100%), and aadA1 (79.6%) were detected from 54 streptomycin resistance strains. sulII (100%) and dhfrXII (100%) were detected from 49 sulfamethoxazole/trimethoprim resistance strains. tetA was detected from 54 (100%) of 54 tetracycline resistance strains. int gene was detected from 54 (100%) of 54 strains. The major drug resistance pattern and resistance gene profile were ampicillin, chloramphenicol, streptomycin, sulfamethoxazole/trimethoprim and tetracycline (ACSSuT) and TEM, cmlA, aadA1, aadA2, strA, strB, sulII, dhfrXII, tetA and int, respectively.

Analysis of virulence gene profiles of Salmonella spp. and Enterococcus faecalis isolated from the freshly slaughtered poultry meats produced in Gyeong-Nam province (경남지역 가금류 도축장 신선육에서 분리한 Salmonella spp.와 Enterococcus faecalis의 독성인자 보유 패턴 분석)

  • Hah, Do-Yun;Cha, Hwi-Geun;Han, Kwon-Seek;Jang, Eun-Hee;Park, Ha-Yeong;Bae, Min-jin;Cho, Ah Reum-Song I;Lee, Hoo-Geun;Ko, Byeong-Hyo;Kim, Do-Kyoung;Hwang, Bo-Won;Kim, Sang-Hyun
    • Korean Journal of Veterinary Service
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    • v.41 no.3
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    • pp.157-163
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    • 2018
  • In order for monitoring of pathogenic bacterial contamination in the freshly slaughtered poultry meats produced in Gyeong-Nam province, we first isolated 4 strains of Salmonella spp. and 32 strains of Enterococcus faecalis from the total 164 samples, then we analyzed potential virulence gene profiles of the bacterial isolates by PCR using species-specific primer. The potential virulence genes we selected in this study were stn, invA, fimA, spvR, and spvC for the isolates of Salmonella spp. and those of esp, cylM, cylA, cylB, gelE, fsrA, fsrB, and fsrC were for the isolates of E. faecalis. The PCR results showed that all 5 virulence genes were detected simultaneously in the all isolates of Salmonella spp. However, there was a diverse occurrence pattern of the virulence genes in the case of E. faecalis. The gene for enterococcal surface protein (esp) was not detected among the isolates (0/32), and the haemolysin gene prevalence rate of cylA, cylB, and cylM were 3.1% (1/32), 9.3% (3/32), and 9.3% (3/32), respectively. Moreover, the genes of gelE, fsrA, fsrB, and fsrC that associated with gelatinase activity were detected in the rate of 53.1% (17/32), 53.1% (17/32), 53.1% (17/32), and 53.1% (17/32), respectively. In conclusion, in the isolates of Salmonella spp., all possessed 5 virulence genes tested, suggesting that they are all related with each other clonally. However, in the case of E. faecalis isolates, the occurrence of the haemolysin genes (cylM, cylA, cylB) and the gelatinase genes (gelE, fsrABC) was highly variable among the isolates.

Comparative Analysis of Salmonella enterica subsp. enterica Serovar Thompson Isolates associated with Outbreaks Using PFGE and wgMLST

  • Youngho Koh;Yunyoung Bae;Min-Jung Lee;Yu-Si Lee;Dong-Hyun Kang;Soon Han Kim
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1605-1614
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    • 2022
  • The strains associated with foodborne Salmonella enterica Thompson outbreaks in Korea have not been identified. Therefore, we characterized S. Thompson strains isolated from chocolate cakes linked to foodborne outbreaks in Korea. A total of 56 strains were isolated from preserved cake products, products in the supply chain distribution, the manufacturer's apparatus, and egg white liquid products used for cream preparation. Subsequently, serological typing, pathogenic gene-targeted polymerase chain reaction (PCR), pulsed-field gel electrophoresis (PFGE), and whole-genome multi-locus sequence typing (wgMLST) were performed to characterize these isolates. The antigen formula of all isolates was 7:k:1,5, namely Salmonella enterica subsp. enterica Serovar Thompson. All 56 isolates harbored invA, his, hin, and stn, and were negative for sefA and spvC based on gene-targeted PCR analyses. Based on PFGE results, these isolates were classified into one group based on the same SP6X01.011 pattern with 100% similarity. We selected 19 strains based on the region and sample type, which were subjected to wgMLST. Although the examined strains showed 100% similarity, they were classified into seven clusters based on allelic differences. According to our findings, the cause of these outbreaks was chocolate cake manufactured with egg white liquid contaminated with the same Salmonella Thompson. Additionally, comparative analysis of wgMLST on domestic isolates of S. Thompson from the three outbreaks showed genetic similarities of over 99.6%. Based on the results, the PFGE and wgMLST combination can provide highly resolved phylogeny and reliable evidence during Salmonella outbreak investigations.

Rapid and Sensitive Detection of Salmonella spp. by Using a Loop-Mediated Isothermal Amplification Assay in Duck Carcass Sample (오리 도체에서 등온유전자증폭기법을 이용한 Salmonella spp. 신속 고감도 검출 기법 연구)

  • Cho, Ae-Ri;Dong, Hee-Jin;Cho, Seongbeom
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.655-663
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    • 2013
  • In this study, a rapid and sensitive detection tool for screening Salmonella spp. by using a loop-mediated isothermal amplification (LAMP) assay targeting the genomic sequence of the invA gene was developed. The inclusivity and exclusivity were both at 100% in the analysis, and the limit of detection (LOD) in a pure S. Enteritidis culture suspended in saline was $3.2{\times}10^3$ CFU/mL at 18.17 min ($R^2$ = 0.9446). The LODs of the LAMP assay in buffered peptone water with Salmonella (BPW) and duck carcass swab sample enriched in BPW with Salmonella (BPWS) after 0 and 12 h incubation were $3.2{\times}10^3$ CFU/mL and $3.2{\times}10^0$ CFU/mL, respectively. Comparing the LODs in BPW with those in BPWS, the LAMP assay was less influenced by compounds of duck carcass swab sample than the PCR assay. Sensitivity and specificity of the LAMP assay in 50 duck carcass swab samples enriched in BPW for 6 h were 96% and 84%, respectively, indicating that the LAMP assay is a rapid, simple and sensitive assay, which can be utilized as a potential screening tool of Salmonella spp. in duck carcass sample.

Serotypes and genotypes of Salmonella isolates from slaughtered pigs (도축돈에서 분리된 살모넬라의 혈청형 및 유전형)

  • Choi, Won-Zong;Jung, Ji-Hun;Won, Ho-Keun;Kang, Zheng-Wu;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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    • pp.1-16
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    • 2008
  • Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.

Molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolated from pigs and cattle (소와 돼지유래 다제내성 Salmonella속 균의 분자유전학적 특성)

  • Lee, Woo-Won;Jung, Byeong-Yeal;Lee, Gang-Rok;Lee, Dong-Soo;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.32 no.1
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    • pp.61-76
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    • 2009
  • At the present study, it was aimed to explore the molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolates from pigs and cattle. A total of 138 Salmonella Typhimurium (S. Typhimurium) isolates were typed with phage, among them, 83.3% of S. Typhimurium tested could divide into a 10 phage types. Definitive type 193 (DT193) (25.4%) and DT195 (24.6%) were exhibited as the dominant types. DT104 and U302 were found from pigs and cattle. On the other hand, S. Enteritidis had 6 phage types, of them, phage type 21 (PT21) and PT11b were the popular types. In the plasmid profiles, 135 of S. Typhimurium isolates were exhibited 1 to 6 plasmid bands which molecular weight ranged from 90 to 2kb. 35 isolates (25.4%) harbored a 90kb plasmid which is thought to be the serotype specific virulence plasmid. Two of twenty five S. Enteritidis had common plasmids at 2 and 1.5kb. With multiplex polymerase chain reaction, virulence genes (invA and spvC) were detected from all Salmonella spp. from 167 of S. Typhimurium, S. Enteritidis and chloramphenicol resistant S. Schwarzengrund, but some drug resistant genes, such as PSE-1, cml/tetR and flo were not determined but other drug resistant genes, for example TEM and int were found. The detection rates of spvC, TEM and int gene was 35.3%, 29.3% and 72.5%, respectively. The TEM gene was highly popular in S. Typhimurium, which was detected from ampicillin and amoxicillin resistant strains as 95.9%. int gene was able to detect from all the isolates identified as multidrug resistsnt (MDR), particularly DT193 was thought as the most prevalent virulence and multidrug resistance isolate. The major plasmid profile and drug resistance pattern of DT193 were 90, 40, 10.5, 6.3, 3.0kb and ACCbDNaPSSuT, respectively. MDR was commonly found in other phage types, particularly DT104, U302 and DT203.

Functional analysis of PEBP2$\alpha$C activity by knockout mouse model (Knockout 마우스 생산에 의한 PEBP2aC 유전자의 생물학적 활성의 규명)

  • ;;;Ito Yoshiaki
    • Proceedings of the Korean Society of Toxicology Conference
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    • 1998.10a
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    • pp.8-13
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    • 1998
  • Polyoma Virus Enhancer core Binding Protein (PEBP2)는 유전자의 전사를 조절하는 hetero-dimeric transcription factor로서 $\alpha$$\beta$ subunit으로 구성되어 있다. $\alpha$ subunit을 coding 하는 유전자중 하나인 PEBP2aB는 급성백혈병과 관련되어있는 t(8;21) 또는 t(12;21)에 의하여 변형됨으로서 백혈병 발병의 원인이 되고 있다 (Miyoshi et al., 1993; .Romana et al., 1995). $\beta$ subunit을 coding 하는 PEBP2$\beta$도 inv(16)에 의하여 변형됨으로서 백혈병을 유도하는 주요 원인이 되고 있다 (Liu et al., 1993). 이 유전자들의 생물학적 활성을 밝히기 위한 연구가 gene targeting에 의한 knockout mouse 생산 방법으로 수행되었다. 그 결과 PEBP2$\alpha$B와 PEBP2$\beta$ 유전자가 definitive hematopoiesis에 있어서 결정적으로 중요한 역할을 하고 있음이 관찰되었다 (Okuda et al., 1996, Wang et al., 1996a, 1996b), 이는 이들 유전자가 bematopoietic master switch 유전자임을 밝힌 중요한 결과로서 이로부터 혈액학 연구 분야의 새로운 장이 열리게 되었다. 또한 이러한 연구 결과들은 PEBP2 family에 속하는 다른 유전자의 생물학적 활성의 연구를 촉진하는 계기가 되었다. 최근 PEBP2$\alpha$A 유전자가 결손된 마우스가 생산되었는데 이 유전자의 경우에는 모든 종류의 뼈의 생성이 완전히 결손됨이 관찰되었다 (Komori et al., 1997). 이는 PEBP2$\alpha$A 유전자가 뼈의 생성을 지배하는 master switch 유전자임을 보여주는 중요한 관찰로서 bone biologist 들의 큰 관심을 모으고 있다. 본 연구팀은 PEBP2 family 유전자 중 유일하게 아직 생물학적 활성이 규명되지 않은 PEBP2$\alpha$C 유전자의 활성을 knockout 마우스를 생산하는 방법에 의하여 분석하였으며 소화기관의 형성에 중요한 역할을 하고 있음을 확인하였다.

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Sugar content and expression of sugar metabolism-related gene in strawberry fruits from various cultivars

  • Lee, Jeongyeo;Kim, Hyun-Bi;Noh, Young-Hee;Min, Sung Ran;Lee, Haeng-Soon;Jung, Jaeeun;Park, Kun-Hyang;Kim, Dae-Soo;Nam, Myeong Hyeon;Kim, Tae Il;Kim, Sun-Ju;Kim, HyeRan
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.90-101
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    • 2018
  • Strawberry (Fragaria ${\times}$ ananassa) is a globally-cultivated and popular fruit crop, prized for its flavor and nutritional value. Sweetness, a key determinant of fruit quality, depends on the sugar composition and concentration. We selected eight strawberry cultivars based on the fruit soluble solids content to represent high and low sugar content groups. The average soluble solid content was $13.6^{\circ}Brix$ (Okmae, Geumsil, Aram, and Maehyang) and $2.9^{\circ}Brix$ (Missionary, Camino Real, Portola, and Gilgyung53), for the high and low sugar content groups, respectively. Sucrose was the main sugar in the cultivars with high sugar content, whereas fructose was the main component in the low sugar content cultivars. Fruit starch concentration ranged from $3.247{\pm}0.056$ to $3.850{\pm}0.055g/100g$, with a 12% higher concentration in the high sugar content cultivars. Additionally, we identified 41 sugar metabolism-related genes in Fragaria ${\times}$ ananassa and analyzed the relationship between their transcripts and the sugar accumulation in fruit. FaGPT1, FaTMT1, FaHXK1, FaPHS1, FaINVA-3, and FacxINV2-1 were highly expressed in the high sugar content cultivars, while FapGlcT, FaTMT2-1, FaPHS2-1, FaSUSY1-1, and FaSUSY1-2 were highly expressed in the low sugar content cultivars. In general, a greater number of genes encoding sugar transporters or involved in sugar synthesis were highly expressed in the high sugar content cultivars. Contrarily, genes involved in sugar degradation were preferentially transcribed in the low sugar content cultivars. Although gene expression was not perfectly proportional to sugar content or concentration, our analysis of the genes involved in sugar metabolism and accumulation in strawberries provides a framework for further studies and for the subsequent engineering of sugar metabolism to enhance fruit quality.