• Title/Summary/Keyword: PCR-dot blot hybridization

Search Result 24, Processing Time 0.019 seconds

Genetic and Phenotypic Diversity of Carbofuran-Degrading Bacteria Isolated from Agricultural Soils

  • Shin, Dong-Hyeon;Kim, Dong-Uk;Seong, Chi-Nam;Song, Hong-Gyu;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
    • /
    • v.22 no.4
    • /
    • pp.448-456
    • /
    • 2012
  • Thirty-seven carbofuran-degrading bacteria were isolated from agricultural soils, and their genetic and phenotypic characteristics were investigated. The isolates were able to utilize carbofuran as a sole source of carbon and energy. Analysis of the 16S rRNA gene sequence indicated that the isolates were related to members of the genera Rhodococcus, Sphingomonas, and Sphingobium, including new types of carbofuran-degrading bacteria, Bosea and Microbacterium. Among the 37 isolates, 15 different chromosomal DNA patterns were obtained by polymerase chain reaction (PCR) amplification of repetitive extragenic palindromic (REP) sequences. Five of the 15 representative isolates were able to degrade carbofuran phenol, fenoxycarb, and carbaryl, in addition to carbofuran. Ten of the 15 representative isolates had 1 to 8 plasmids. Among the 10 plasmid-containing isolates, plasmid-cured strains were obtained from 5 strains. The cured strains could not degrade carbofuran and other pesticides anymore, suggesting that the carbofuran degradative genes were on the plasmid DNAs in these strains. When analyzed with PCR amplification and dot-blot hybridization using the primers targeting for the previously reported carbofuran hydrolase gene (mcd), all of the isolates did not show any positive signals, suggesting that their carbofuran hydrolase genes had no significant sequence homology with the mcd gene.

Molecular pathological interactions between Apple stem grooving virus (ASGV) and its fungi.

  • Hyekyung Shim;Lee, Hyunjeong;Seungbeom Hong;Park, Dae-Sup;DaeRobert A Samson;Hyeongjin Jee;Lee, Sukchan
    • Proceedings of the Korean Society of Plant Pathology Conference
    • /
    • 2003.10a
    • /
    • pp.122-123
    • /
    • 2003
  • Apple stem grooving virus (ASGV) belongs to Capillovirus and infects pome fruits. Transmission mode of ASGV is known by grafting and mechanical inoculation into susceptible hosts, not by any other natural vectors. But we have observed the spread of ASGV in the field without mechanical inoculation or grafting. Transmission seems to be occurred from tree-to-tree and tree-to-susceptible herbaceous plants along but not across ditches in the field. In order to ascertain this possibility, various fungi were isolated and cultured from ASGV-infected plants and 69 isolates were characterized. By means of RNA dot-blot hybridization and PCR analysis, 3 isolates were sorted out for further studies. The isolates were identified to Tataromyces sp. and belonged to Phenicillium by morphological characteristics and molecular markers. As an experimental host, 10 kidney beans (Phaseolus vulgaris) were screened and Kyunggi-5 was selected for virus amplification and symptom development. Kyunggj-5 infected by fungi which seemed to carry ASGV showed the typical disease symptoms and viral coat protein genes were detected from all tested plants. To confirm the Koch's rule, fungi cultured from inoculation origins of kidney bean were grown on PDA media and re-inoculated to hosts. The fungi isolated from inoculation origins induced the typical disease symptoms on hosts. However virus free fungi did not induce any symptom on the experimental hosts. This bioassay showed that these typical symptoms were caused by virus, not fungi.

  • PDF

Human Papillomavirus (HPV) Infection in Women Participating in Cervical Cancer Screening from 2006 to 2010 in Shenzhen City, South China

  • Wang, Yue-Yun;Li, Li;Wei, Sheng;Peng, Ji;Yuan, Shi-Xin;Xie, Jian-Sheng;Liu, Zhi-Hua
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.12
    • /
    • pp.7483-7487
    • /
    • 2013
  • Purpose: Human papillomavirus (HPV) infection plays an important role in the development of cervical cancer, but the prevalence of HPV infection in women of Shenzhen city remains unclear. The present study was performed to describe the change of cervical HPV infection in females who participated in voluntary cervical cancer screening from 2006 to 2010 in Shenzhen city, China. Methods: A total of 4, 413 women were recruited. HPV infections were genotyped by polymerase chain reaction (PCR) and reversed dot blot hybridization in Shenzhen Maternity and Child Health Hospital. Results: The prevalence of HPV infection was 13.8%. The five most commonly found HPV types were HPV16 (3.47%), HPV58 (1.68%), HPV33 (1.38%), HPV43 (1.36%) and HPV18 (1.27%). The secular trends of major HPV type-specific were diverse. Among of them, the prevalence of HPV18 increased sharply while others increased slowly or even decreased in the period. The change of total HPV, single HPV and multiple HPV infection were similar during the five years. Conclusions: Our findings suggested that HPV infection is common with HPV16 and HPV 58 as the primary subtypes in women in Shenzhen city.The prevalence of HPV 18 infection is increasing faster than any others, which will lead it to be one of the main subtypes in this city in the future.

The Prevalence of Oral Spirochetes in Korean Adult Periodontitis (한국인 성인성 치주염 환자에서의 구강 스피로헤타의 분포)

  • Kim, Hay-Hyun;Choi, Bong-Kiu;Choi, Seong-Ho;Chai, Jung-Kiu;Kim, Chong-Kwan;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
    • /
    • v.28 no.4
    • /
    • pp.659-678
    • /
    • 1998
  • In the present study, oligonucleotide probes based on 16S rRNA were taken to investigate the diversity of oral spirochetes without culture method. This is the first study that revealed oral spirochetes of both presently cultivable and uncultured oral spirochetes in Korean adult periodontitis patients. Subgingival plaque samples were taken from diseased sites(probing depth ${\geq}6\;mm$, experimental group, n=116) and healthy sites(probing depth${\leq}3mm$, control 1 group, n=28) in 29 patients with adult periodontitis, and from 20 periodontally healthy subjects(probing depth${\leq}3mm$, control 2 group, n=100). Following being examined under phase-contrast microscope, all samples were submitted to dot-blot hybridization after polymerase chain reacton with eubacterial primers. 5 species-specific probes(TVIN, TDEN, TMAL, TSOC, and TPEC) and 7 group-specific probes(TRE I, TRE II, TRE III, TRE IV, TRE V, TRE VI, and TRE VII) were used one by one for the identification of both cultivable and so far uncultivable oral spirochetes. All probes were labeled with digoxigenin(DIG)-ddUTP and detected by chemilumininescence. The following results were obtained. 1. Under phase-contrast microscope, 91.37% and 14.28% of oral spirochetes were observed in the experimental and control 1 groups, respectively. None of oral spirochetes were observed in control 2 group. 2. With universal probe, 98.27%, 46.42%, and 22.0% of oral spirochetes were observed in experimental, control 1, and control 2 groups, respectively. 3. With specific probe, 95.68%, 35.71%, and 19.0% of oral spirochetes were observed in experimental, control 1, and control 2 groups, respectively. 4. With species-specific probes, T. socranskii were recovered in a high percentage of sites(81.89%) examined, followed by T. maltophilum(50.0%), T. vincentii(36.20%), T. denticola(13.79%), respectively. With group- specific probes, TRE IV was recovered in a high percentage of sites(85.34%) examined, followed by TRE II(77.58%), TRE I(56.89%), TRE III(25.86%), TRE VI(5.17%), and TRE V(2.58%), respectively. 5. T. vincentii were only observed in the diseased sites, not in the healthy sites. 6. Neither T. pectinovorum nor group VII oral spirochetes were observed in any sites. The findings warrant further investgations of the recovered spirochetes to elucidate the possible associations of oral spirochetal prevalence in race and types of periodontitis, pathogenesis of T. vincentii and the possible distributional change of oral spirochetes before and after treatments.

  • PDF