• 제목/요약/키워드: Primers

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Genetic Differences within and between Populations of Korean Catfish (S. asotus) and Bullhead (P. fulvidraco) Analysed by RAPD-PCR

  • Yoon, Jong-Man;Kim, Jong-Yeon
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2004년도 수산관련학회 공동학술대회 발표요지집
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    • pp.321-322
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    • 2004
  • Of the 20 arbitrarily chosen primers, six oligonucleotides decamer primers were used on the basis of the number of the polymorphisms generated in catfish (Silurus asotus) from Yesan and bullhead (Pseudobagrus fulvidraco) from Dangjin in Korea. Six primers were used generating a total of 602 scorable bands in catfish and 195 in bullhead population, respectively, ranging in size of DNA fragments from less than approximately 100 to larger than 2,000 base pairs (bp). Six primers yielded 199 polymorphic fragments (33.1 %) in catfish and 47 (24 %) in bullhead, respectively. In the present study, a total of 328 common fragments (an average of 54.7 per prime.) were observed in catfish population, whereas 84 (an average of 14.0 per prime.) in bullhead. The total number of specific fragments in catfish and bullhead population were 76and 64, respectively.

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비스페놀A 처리 수컷 잉어에서 비텔로제닌 mRNA의 발현 (Vitellogenin mRNA Induction in Male Carp Treated with Bisphenol A)

  • 박응로;최필선;이철우;류지성;남성숙;류홍일;전성환;나진균;박광식
    • Toxicological Research
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    • 제17권3호
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    • pp.181-186
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    • 2001
  • The estrogenic potency of bisphenol A using reverse trancriptase-PCR response of liver vitellogenin mRNA in male carp was studied. For this, six combination of primers which were synthesized on the basis of cDNA consensus region of various species, were evaluated and one pair of primers was selected as the best to show 286 bp size-transcript. By using the selected primers, vitellogenin mRNA induction in carp treated with bisphenol A was measured and the chemical showed dose-and time-dependent Induction response. From this result, it was concluded that RT-PCR technique wing the selected primers in this study can be wed to monitor the estrogenic effects exerted In carp living in Korean freshwater.

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Genetic variation of BIV isolates characterized by PCR using degenerate primers

  • Kwon, Oh-Sik;Sninsky, John J.
    • Journal of Microbiology
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    • 제33권3호
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    • pp.252-259
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    • 1995
  • The PCR was employed to detect and characterize the bovine immunodeficiency-like virus (BIV), which is a newly recognized member of the I entivirinae of the retroviruses. Degenerate primers representing the conserved regions in the pol genes of the Lentivirinae, were used to detect proviral DNA obtained from the bovine embryonic spleen cell cultures infected with BIV. The PCR amplified DNA fragment was molecularly cloned and sequenced. The BIV DNA fragment contained a sequence identical to that reported by Garvey et al. (Garvey et al., 1990. Virology, 175, 391-409). With the degenerate primers, peripheral blood mononuclear cells (PBMCs) of sick cattle and cells cultured with BIV were tested to determine genetic variation of BIV pol conserved sequence. We found the sequence heterogeneity within cultures and most variations occurred at the third base of codons that would not lead to amino acid substitutions. Another change was GAG (Glu) to AAG (Lys) within the BIV isolates. Interestingly, the altered sequence is also found in other lentiviruses such as HIV-2, SIV mac, CAEV and EIAV.

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Development of Penicillium italicum-Specific Primers for Rapid Detection among Fungal Isolates in Citrus

  • Chen, Kai;Tian, Zhonghuan;Jiang, Fatang;Long, Chao-an
    • Journal of Microbiology and Biotechnology
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    • 제29권6호
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    • pp.984-988
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    • 2019
  • Blue mold in citrus is caused by Penicillium italicum. In this study, the P. italicum-specific primers were developed for rapid detection based on the conserved genes RPB1 and RPB2 among Penicillium genomes. The two primer pairs RPB1-a and RPB1-b proved to be specific to detect P. italicum. The PCR assay among 39 fungal isolates and the colonial, pathogenic morphologies and molecular methods validated the specificity and reliability of these two primer pairs. This report provided a method and P. italicum-specific primers, which might greatly contribute to citrus postharvest industry.

PCR-RAPD를 이용한 제주말의 유전적 다양성분석 (Genetic Diversity Analysis of the Cheju Horse Using Random Amplified Polymorphic DNAs)

  • Cho, Byung-Wook;Lee, Kil-Wang
    • 생명과학회지
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    • 제14권3호
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    • pp.521-524
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    • 2004
  • 본 연구는 short oligonucleotide primer를 이용하여 마 품종간 유전 분석을 실시 하고자 PCR증폭 기법을 확립하고, 확립된 기술을 이용하여 제주도에 사육중인 천념기념물 347호로 등록된 제주말과 경주마로 잘 알려진 더러브렛간의 유전적인 다양성을 분석한 결과 마 품종간 차이를 보이는 DNA marker는 9개의 primer에서 확인되었으며, 이중 6개의 primer에서 더러브렛 특이 밴드와 나머지 3개에서 제주 마 특이 RAPD 밴드가 확인되어 cloning과 sequencing후에 SCAR primer를 제작하여 마 품종 식별에 활용할 수 있을 것으로 사료되며, 본 연구결과 RAPD표지인자는 마 품종간의 유전 분석에 매우 유용한 것으로 판단되었다.

레진 표면 처리제의 도포에 따른 의치상 레진과 개상용 레진의 결합강도에 관한 연구 (A STUDY ON THE BOND STRENGTH OF REBASE RESIN TO DENTURE BASE RESIN BY APPLICATION OF PRIMERS)

  • 문병준;방몽숙
    • 대한치과보철학회지
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    • 제36권1호
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    • pp.50-63
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    • 1998
  • This study investigated the effects of resin surface primers for rebase resins on the surface texture of denture base resins by the use of scanning electron microscopy. This study also evaluated the bond strength of rebase resins to denture base resin. The denture base resin in this study was Vertex RS (Dentimex Co., Ltd., Holland). The rebase resins used were Tokuso Rebase (Tokuyama Co., Ltd., Japan), Metabase (Sun Medical Co., Ltd., Japan), New True Liner(Harry J, Bosworth Co., Ill.), and Reverse (Nissin Co., Ltd., Japan). The test samples were divided into four parts: Group 1 : Treated with primer with brush. Group 2 : Immersed in the primer for 5 seconds. Group 3 : Immersed in the primer for 10 seconds. Group 4 : Immersed in the primer for 30 seconds. Control group : not treated with primer The results were as follows; 1. The bond strength of rebase resins to denture base resin is increased by application of primers. 2. Regardless of the rebase resin type, there was no significant difference among the bond strength in groups G1, G2, G3, G4. There was a significant difference with the control. (P<0.05) 3. Regardless of each group, the bond strength according to the rebase resin type was decreased in the following order: Tokuso Rebase, New Tru Liner, Reverse and then Metabase. 4. Under the scanning electron microscope, brush application produced a softening of the denture base surface. After immersion, all primers produced a spongelike structure on the denture base resin surface. The results of this study suggest that primers produce a significant effect by means of brush application, therefore it is recommended as the most convenient and logical procedure for application of the primers.

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tufA gene as molecular marker for freshwater Chlorophyceae

  • Vieira, Helena Henriques;Bagatini, Inessa Lacativa;Guinart, Carla Marques;Vieira, Armando Augusto Henriques
    • ALGAE
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    • 제31권2호
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    • pp.155-165
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    • 2016
  • Green microalgae from the class Chlorophyceae represent a major biodiversity component of eukaryotic algae in continental water. Identification and classification of this group through morphology is a hard task, since it may present cryptic species and phenotypic plasticity. Despite the increasing use of molecular methods for identification of microorganisms, no single standard barcode marker is yet established for this important group of green microalgae. Some available studies present results with a limited number of chlorophycean genera or using markers that require many different primers for different groups within the class. Thus, we aimed to find a single marker easily amplified and with wide coverage within Chlorophyceae using only one pair of primers. Here, we tested the universality of primers for different genes (tufA, ITS, rbcL, and UCP4) in 22 strains, comprising 18 different species from different orders of Chlorophyceae. The ITS primers sequenced only 3 strains and the UCP primer failed to amplify any strain. We tested two pairs of primers for rbcL and the best pair provided sequences for 10 strains whereas the second one provided sequences for only 7 strains. The pair of primers for the tufA gene presented good results for Chlorophyceae, successfully sequencing 21 strains and recovering the expected phylogeny relationships within the class. Thus, the tufA marker stands out as a good choice to be used as molecular marker for the class.

RAPD를 이용한 고추(Capsicum annuum) 유전자원의 분류 (Classification of Capsicum annuum Germplasm Using Random Amplified Polymorphic DNA)

  • 남승현;최근원;유일웅
    • 원예과학기술지
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    • 제16권4호
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    • pp.503-507
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    • 1998
  • 본 연구는 RAPD표지를 이용하여 국내외에서 수집된 고추 유전자원들간의 유전적관계를 평가하고자 수행되었다. Random primer를 이용한 고추의 PCR반응은 $MgCl_2$ 3mM, Taq. DNA polymerase 1.5U, 주형 DNA 10ng, dNTPs $200{\mu}M$, random primer 200nM 그리고 $42^{\circ}C$의 annealing 온도조건으로 최적화하였다. 80개의 random primer로부터 높은 밴드선명도와 재현성을 보이는 16개가 선발되었으며 70%의 GC함량을 갖는 primer들이 GC함량이 60%인 것보다 DNA증폭에 있어 효과적이었다. 31개의 고추품종및 계통들에 대해 71개의 polymorphic밴드와 22개의 monomorphic밴드를 포함하는 총 93개의 DNA밴드가 선발된 16개의 random primer들로부터 형성되었다. Primer당 약 4.4개의 polymorphic밴드가 형성되었다. 이들 71개의 polymorphic밴드를 이용하여 유사도지수가 구해졌으며 이를 근거로 31고추 계통 또는 품종들을 뚜렷이 구분하는 dendrogram이 작성되었다.

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Molecular Characterization of Rathi and Tharparkar Indigenous Cattle (Bos indicus) Breeds by RAPD-PCR

  • Sharma, Amit Kumar;Bhushan, Bharat;Kumar, Sanjeev;Kumar, Pushpendra;Sharma, Arjava;Kumar, Satish
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권9호
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    • pp.1204-1209
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    • 2004
  • Random amplification of polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out using DNA samples of 30 animals of Rathi cattle and 42 animals of Tharparkar cattle. Genomic DNA was isolated as per standard protocol and evaluated for its quality, purity and concentration. Twenty three random primers were screened out of which 15 primers yielded satisfactory amplifications and were used for further analysis. Average numbers of polymorphic fragments per primer were 7.07${\pm}$0.86 in Rathi and 6.80${\pm}$0.61 in Tharparkar cattle. The percentage of polymorphic bands in these two cattle breeds were 86 and 87%, respectively. Within breed genetic similarities for pooled over primers in the animals of Rathi and Tharparkar breeds were .577${\pm}$0.30 and 0.531${\pm}$0.02, respectively on the basis of band frequency (BF) and 0.645${\pm}$0.04 and 0.534${\pm}$0.04, respectively on the basis of band sharing (BS). Averages of between breed genetic similarities for pooled over primers were 0.97 and 0.92 according to BF and BS, respectively, which reflect higher degree of genetic similarity between Rathi and Tharparkar cattle breeds. Index of genetic distance based on BF and BS for pooled over primers was 0.030${\pm}$0.011 and 0.088${\pm}$0.031, respectively. Percentage of polymorphic bands and within-breed genetic similarities on the basis of band frequency (BF) and band sharing (BS) for pooled over primers revealed higher genetic similarity in Rathi than Tharparkar cattle population. High estimates of between breed genetic similarities for pooled over primers indicated that either Rathi is having decent from Tharparkar or both the cattle breeds are having common descent. Low value of Index of genetic distances between these two cattle breeds may be due to the fact that Rathi and Tharparkar cattle breeds are the native of Thar Desert in Northwest India. The results of between breed genetic distances also confirm the existence of high degree of genetic similarity between these two breeds of cattle.

16S rDNA sequence에 대한 종특이성 primer를 이용한 중합효소연쇄반응증폭에 의한 Porphyromonas endodontalis의 동정에 관한 연구 (A STUDY ON THE IDENTIFICATION OF Porphyromonas endodontalis BY PCR USING SPECIES SPECIFIC PRIMERS FOR THE 16S rDNA)

  • 엄승희;임성삼;배광식
    • Restorative Dentistry and Endodontics
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    • 제24권1호
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    • pp.13-25
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    • 1999
  • P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

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