• Title/Summary/Keyword: Intraspecies

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Quantitative analysis of mitochondrial DNA in porcine-mouse cloned embryos

  • Hyeonyeong Shin;Soyeon Kim;Myungyoun Kim;Jaeeun Lee;Dongil Jin
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.767-778
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    • 2023
  • The aim of the research is to identify that porcine oocytes can function as recipients for interspecies cloning and have the ability to develop to blastocysts. Furthermore each mitochondrial DNA (mtDNA) in interspecises cloned embryos was analyzed. For the study, mouse-porcine and porcine-porcine cloned embryos were produced with mouse fetal fibroblasts (MFF) and porcine fetal fibroblasts (PFF), respectively, introduced as donor cells into enucleated porcine oocytes. The developmental rate and cell numbers of blastocysts between intraspecies porcine-porcine and interspecies mouse-porcine cloned embryos were compared and real-time polymerase chain reaction (PCR) was performed for the estimate of mouse and porcine mtDNA copy number in mouse-porcine cloned embryos at different stages.There was no significant difference in the developmental rate or total blastocyst number between mouse-porcine cloned embryos and porcine-porcine cloned embryos (11.1 ± 0.9%, 25 ± 3.5 vs. 10.1 ± 1.2%, 24 ± 6.3). In mouse-porcine reconstructed embryos, the copy numbers of mouse somatic cell-derived mtDNA decreased between the 1-cell and blastocyst stages, whereas the copy number of porcine oocyte-derived mtDNA significantly increased during this period, as assessed by real-time PCR analysis. In our real-time PCR analysis, we improved the standard curve construction-based method to analyze the level of mtDNA between mouse donor cells and porcine oocytes using the copy number of mouse beta-actin DNA as a standard. Our findings suggest that mouse-porcine cloned embryos have the ability to develop to blastocysts in vitro and exhibit mitochondrial heteroplasmy from the 1-cell to blastocyst stages and the mouse-derived mitochondria can be gradually replaced with those of the porcine oocyte in the early developmental stages of mouse-porcine cloned embryos.

Functional Identification and Genetic Analysis of O-Antigen Gene Clusters of Food-Borne Pathogen Yersinia enterocolitica O:10 and Other Uncommon Serotypes, Further Revealing Their Virulence Profiles

  • Bin Hu;Jing Wang;Linxing Li;Qin Wang;Jingliang Qin;Yingxin Chi;Junxiang Yan;Wenkui Sun;Boyang Cao;Xi Guo
    • Journal of Microbiology and Biotechnology
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    • v.34 no.8
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    • pp.1599-1608
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    • 2024
  • Yersinia enterocolitica is a globally distributed food-borne gastrointestinal pathogen. The O-antigen variation-determined serotype is an important characteristic of Y. enterocolitica, allowing intraspecies classification for diagnosis and epidemiology purposes. Among the 11 serotypes associated with human yersiniosis, O:3, O:5,27, O:8, and O:9 are the most prevalent, and their O-antigen gene clusters have been well defined. In addition to the O-antigen, several virulence factors are involved in infection and pathogenesis of Y. enterocolitica strains, and these are closely related to their biotypes, reflecting pathogenic properties. In this study, we identified the O-AGC of a Y. enterocolitica strain WL-21 of serotype O:10, and confirmed its functionality in O-antigen synthesis. Furthermore, we analyzed in silico the putative O-AGCs of uncommon serotypes, and found that the O-AGCs of Y. enterocolitica were divided into two genetic patterns: (1) O-AGC within the hemH-gsk locus, possibly synthesizing the O-antigen via the Wzx/Wzy dependent pathway, and (2) O-AGC within the dcuC-galU-galF locus, very likely assembling the O-antigen via the ABC transporter dependent pathway. By screening the virulence genes against genomes from GenBank, we discovered that strains representing different serotypes were grouped according to different virulence gene profiles, indicating strong links between serotypes and virulence markers and implying an interaction between them and the synergistic effect in pathogenicity. Our study provides a framework for further research on the origin and evolution of O-AGCs from Y. enterocolitica, as well as on differences in virulent mechanisms among distinct serotypes.

Sex Pheromone Trapping of Spodoptera frugiperda (Noctuidae: Lepidoptera) in Korea and the Distribution of Intraspecies-specific Single Nucleotide Polymorphisms in the Cytochrome c Oxidase Subunit 1 (CO1) (성페로몬을 이용한 열대거세미나방 포획과 시토크롬 옥시다제 1(CO1)에서 종내 변이군 특이적 단일염기다형성 분포)

  • Seo, Bo Yoon;Jung, Jin Kyo;Lee, Gwan-Seok;Yang, Chang Yeol;Cho, Jumrae;Kim, Yang Pyo
    • Korean journal of applied entomology
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    • v.59 no.3
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    • pp.217-231
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    • 2020
  • In 2019, the sex pheromones of Spodoptera frugiperda were used to examine moth trapping in cornfields in Gochang, Korea. Four types of traps were prepared, two funnel-types and two delta-types, each baited with 300 or 1000 ㎍ of a two-component (2C) blend of synthetic sex pheromones [100% (Z)-9-tetradecenyl acetate (Z9-14Ac) and 2% (Z)-7-dodecenyl acetate (Z7-12Ac)]. The greatest number of S. frugiperda were captured in the 300 ㎍ funnel-type trap (first catch: August 6). Large numbers of Mythimna loreyi (a non-target) were also caught in the funnel-type traps. Two wing-type traps were baited with 1000 ㎍ of the 2C blend or a four-component (4C) blend [100% (Z)-9-tetradecenyl acetate, 8% (Z)-11-hexadecenyl acetate (Z11-16Ac), 2% (Z)-7-dodecenyl acetate, and 1% (Z)-9-dodecenyl acetate (Z9-12Ac)] and the capture efficiency was assessed. Low numbers of S. frugiperda were captured regardless of the blend, and more M. loreyi were captured using the 4C blend. The two intraspecies groups clustered separately in a phylogenetic tree constructed using partial sequences (1004 bp) of cytochrome c oxidase subunit 1 (CO1). Of the 70 S. frugiperda captured in the pheromone traps, 66 belonged to CO1-RS (CO1 rice-strain) and 4 to CO1-CS (CO1 corn-strain). Twelve consistent single nucleotide polymorphisms (SNPs) were identified in CO1 between the CO1-RS and CO1-CS groups of S. frugiperda. Of the 73 S. frugiperda, 4 had the same SNP pattern as the CO1-CS group (including the corn strain) and 69 had the same SNP pattern as the CO1-RS group (including the rice strain).

Identification of Mycobacterium avium complex (MAC) Clinical Strains to a Species Level by Sequencing and PCR-SSCP Analysis of rpoB DNA (비결핵항산성균의 rpoB DNA 염기서열과 SSCP pattern 분석에 따른 Mycobacterium avium complex (MAC) 임상분리균주의 동정)

  • Kim, Bum-Joon;Lee, Seung-Hyun;Lee, Kuen-Hwa;Park, Chung-Kyu;Choi, Myung-Sik;Kim, Ik-Sang;Choi, Sung-Bai;Hwang, Eung-Su;Cha, Chang-Yong;Kim, Sang-Jae;Bai, Gill-Han;Kook, Yoon-Hoh
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.491-500
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    • 1999
  • A recent study showed that comparative sequence analysis of rpoB DNAs could reveal natural relationships in genus Mycobacterium [J Clin Microbial. 37 (6). 1999]. rpoB DNAs showed interspecies variation and intraspecies conservation. Based on these data, we developed polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) protocols which enable species differentiation in genus Mycobacterium. When this assay was applied to 24 clinical isolates identified as M. avium complex (MAC) by biochemical test, these were successfully differentiated into M. avium and M. intracellulare. These results were concordant with those obtained by 16s rDNA analysis. It is the first report that PCR-SSCP analysis of rpoB DNA could be used for species differentiation of MAC strains.

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Comparisons of Isozyme Patterns in Pythium Species and Application to Pythium Systematics (Isozyme을 이용한 Pythium species의 비교 및 Pythium systematics에의 이용)

  • Lee, Youn-Su
    • The Korean Journal of Mycology
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    • v.21 no.4
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    • pp.293-300
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    • 1993
  • For the enzymes AAT, GmDH, ME, GPI, LDH and IDH, nine, seven, four, nine, seven, and four different phenotypes, respectively, were observed. All six isolates of an unidentified sterile Pythium sp. isolated from field soil showed the same band positions for all six enzymes compared. These phenotypes were not similar to any of the known Pythium species. Two isolates of unknown Pythium species (145 and 299) showed the same band positions for all six enzymes. The phenotypes for all three unknown Pythium spp. were different from the other species in the experiment. Five isolates of P. heterothallicum showed the same band positions for all enzymes compared except one enzyme, lDH. Two isolates of P. torulosum showed the same band petitions for enzymes AAT, GmDH and ME, and three isolates of P. totulosum showed the same positions for enzymes GPI, LDH, and IDH. Single isolates of P. spinosum and P. irregulare showed the same band positions for enzymes AAT, GmDH and GPI. In conclusion, sterile types of Pythium species showed 100% similarities among themselves but did not show any similarity with all isolates of P. heterothallicum and P. spinosum isolate, and showed very low similarities with other isolates in general except with unknown Pythium isolate 306. Similarity levels between different species were low in general with few exceptions.

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Effects of Incompatibility on Protoplast Fusion between intra-and inter Species in Basidiomycete, Pleurotus spp. (느타리버섯의 불화합성(不和合性)이 종내(種內) 및 종간(種間) 원형질체(原形質體) 융합(融合)에 미치는 영향(影響))

  • Go, Seung-Joo;You, Chang-Hyun;Shin, Gwan-Chull
    • The Korean Journal of Mycology
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    • v.17 no.3
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    • pp.137-144
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    • 1989
  • Effects of incompatibility existing between intra-and interspecies in Pleurotus spp. on protoplast fusion, clamp formation of their fusants and fruitbody production were investigated. Protoplast fusion between intra-and interspecies of the fungus was achieved by Poly ethylene glycol treatment. The fusion frequency between intraspecies was a little higher than that of interspecies. Fusion frequency between interspecies was not correlated with their similarities based on isozyme patterns. In case of protoplast fusion between intra-and interspecies, the fusants from the compatible isolates produced normal fruit bodies, while those from the incompatible isolates did not produce clamp connections and fruit bodies except those of a few isolates presumed mutants.

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Establishment of the Korean Tolerable Daily Intake of Bisphenol A Based on Risk Assessments by an Expert Committee

  • Choi, Chan-Woong;Jeong, Ji-Yoon;Hwang, Myung-Sil;Jung, Ki-Kyung;Lee, Kwang-Ho;Lee, Hyo-Min
    • Toxicological Research
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    • v.26 no.4
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    • pp.285-291
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    • 2010
  • Recently, reproductive and neurobehavioral effects of bisphenol A (BPA) have been documented, and thus a review was requested for BPA management direction by the government. Therefore, this study was performed to establish a Korean tolerable daily intake (TDI) for BPA. An expert committee, consisting of specialists in fields such as toxicology, medicine, pharmacology, and statistics, was asked to evaluate BPA health based guidance values (HbGVs). Although many toxicological studies were reviewed to select a point of departure (POD) for TDI, rat and mouse reproductive studies by Tyl et al. (2002, 2006), which were performed according to GLP standards and OECD guidelines, were selected. This POD was the lowest value determined from the most sensitive toxicological test. The POD, a NOAEL of 5 mg/kg bw/day, was selected based on its systemic toxicity as critical effects. An uncertainty factor of 100 including interspecies and intraspecies differences was applied to calculate the TDI. According to the evaluation results, a TDI of BPA for Korean was suggested at 0.05 mg/kg bw/day. In addition, the BPA exposure level based on food consumption by the Korean population was estimated as 1.509 ${\mu}g/kg$ bw/day, and the HI was evaluated at 0.03 when the TDI of 0.05 mg/kg bw/day was applied. This HI value of 0.03 indicated that hazardous effects would not be expected from BPA oral exposures. Although highly uncertain, further studies on low dose neurobehavioral effects of BPA should be performed. In addition, it is recommended that the 'as low as reasonably achievable' (ALARA) principle be applied for BPA exposure from food packaging materials in newborn infants and children.

Genetic Diversity and Population Structure of Brassica juncea by Random Amplified Polymorphic DNA (RAPD) (RAPD를 이용한 겨자의 유전적 다양성과 집단구조)

  • Oh, Yung-Hee;Moon, Sung-Gi;Chae, Yang-Hee;Hong, Hwa-Jin;Cho, Min-Cheol;Park, So-Hye;Huh, Man-Kyu
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1538-1543
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    • 2010
  • This study was conducted to examine the genetic diversity and population structure of 17 Brassica juncea populations in Korea. The technique of random amplified polymorphic DNA (RAPD) produced 60 polymorphic loci and 18 monomorphic loci. In a simple measure of intraspecies variability by the percentage of polymorphic bands, the Jindo population of Cheonnam showed the highest (29.5%). The cultivar exhibited the lowest variation (12.8%). Mean number of alleles per locus (A) and the effective number of alleles per locus ($A_E$) were 1.221 and 1.167, respectively. As the typical populations of this species were small, isolated, and patchily distributed in their natural populations, they maintained a low level of genetic diversity of fourteen primers. On a per locus basis, total genetic diversity values ($H_T$) and interlocus variation in the within-population genetic diversity ($H_S$) were 0.347 and 0.141, respectively. On a per-locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) was 0.589. This indicated that about 58.9% of the total variation was among populations. The estimate of gene flow, based on $G_{ST}$, was very low among Korean populations of B. juncea ($N_m$=0.617). These results suggest that the geological distance dispersal of wild B. juncea is the best event. RAPD markers are very effective in classifying natural population levels of B. juncea in Korea.

Phylogenetic Analysis of the Genus Gliocladium and its Related Taxa by Comparing the Sequences of Internal Transcribed Spacers and 5.8S r-DNA (Ribosomal DNA의 Internal Transcribed Spacer(ITS) 부위의 염기서열 분석에 의한 Gliocladium 속과 근연속에 관한 계통 분류학적 연구)

  • Park, Ju-Young;Kim, Gi-Young;Ha, Myoung-Gyu;Shin, Young-Kook;Park, Yong-Ha;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.27 no.3 s.90
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    • pp.191-197
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    • 1999
  • The phylogenetic position of Gliocladium and its related taxa were investigated, using the neighbor-joining method of the sequences from internal transcribed spacers (ITS1 and ITS2) and 5.8S ribosomal DNA (rDNA). It was focused especially on the generic concept by comparing with the related genera such as Trichoderma, Hypocrea, Verticillium, Penicillium and Talaromyces. Gliocladium species and its related genus were divided into three groups by the phylogenetic analysis using the neighbor-joining method. The first group includes Penicillium-like strains such as Penicillium, Tararomyces, Verticillium and one species of Gliocladium (G. cibotii JCM 9203 and JCM 9206). Especially, Gliocladium cibotii JCM 9203 is thought to be the similar species with Verticillium bulbillosum JCM 9214. Between these two species, Gliocladium cibotii and Verticillium bulbillosum, the intraspecies concept needs to examined with culture condition. and morphological properties. The second group includes two species Verticillium, Verticillium tricorpus and Verticillium albo-atrum which extracted from the GenBank database in NCBI (National Center for Biotechnology Information). Trichoderma-like strains, such as Trichoderma, Hypocrea and several species of Gliocladium are included in the third group. Also, Gliocladium penicillioides IFO 5869 and Gliocladium catenulatum ATCC 10523 formed the subgroup of Trichoderma-like strains. The species of Gliocladium were dispersed in Trichoderma-like and Penicillinum-like group, and only one species of Gliocladium cihotii used in our study was located in Penicillium-like genus group. The species of Verticillium appeared in all three groups and the species of Trichoderma formed the monophylogeny with Hypocrea (telemorph). Also, Gliocladium virens was grouped with Trichoderma harzianum with a high bootstrap value, supporting that Gliocladium virens is to be placed in Trichoderma. The results suggest that Gliocladium is polyphyletic, and is more Trichoderma-like than Penicillium-like.

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The Frequency of Detecting Prevotella intermedia and Prevotella nigrescens in Korean Adult Periodontitis Patients (한국인 치주 감염 환자에서의 Prevotella intermedia와 Prevotella nigrescens의 발현빈도)

  • Peck, Seung-Yup;Ku, Young;Rhyu, In-Cheol;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.419-429
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    • 2000
  • Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

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