• 제목/요약/키워드: Restriction Fragment Length

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소 바이러스성 설사병 바이러스의 유전자 재조합 DNA clone의 작성에 관한 연구 (Construction of recombinant DNA clone for bovine viral diarrhea virus)

  • 여상건
    • 대한수의학회지
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    • 제32권3호
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    • pp.389-398
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    • 1992
  • Molecular cloning was carried out on the Danish strain of bovine viral diarrhea virus(BVDV) to construct strategy for the diagnostic tools and effective vaccine of BVD afterwards. A recombinant DNA clone(No. 29) was established successfully from cDNA for viral RNA tailed with adenine homopolymer at 3'-end. $^{32}P$-labeled DNA probes of 300~1,800bp fragments, originating from the clone 29, directed specific DNA-RNA hybridization results with BVDV RNA. Recombinant DNA of the clone 29 was about 5,200bp representing 41.6% of the full length of Danish strain's RNA, and restriction sites were recognized for EcoR I, Sst I, Hin d III and Pst I restriction enzymes in the DNA fragment.

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Muscle-Specific Creatine Kinase Gene Polymorphisms in Korean Elite Athletes

  • Kang, Byung-Yong;Kang, Chin-Yang;Lee, Kang-Oh
    • Toxicological Research
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    • 제19권2호
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    • pp.115-121
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    • 2003
  • In view of the importance of muscle-specific creatine kinase (CKMM) gene as a genetic factor for athletic performance, we investigate the relationship between elite athletic performance and two restriction fragment length polymorphisms (Ncol and Taql RFLPs) in the CKMM gene. Genomic DNA was extracted from white blood cells of 98 unrelated male Korean elite athletes and 04 sedentary controls, respectively. Two genetic polymorphisms in the CKMM gene were detected by the polymerase chain reaction and the digestion with restriction endonucleases, Ncol and Taql, respectively. There were no significant associations between two genetic polymorphisms in the CKMM gene and elite athletic performance or clinical parameters in our subjects. Therefore, these findings suggest that two genetic polymorphisms in the CKMM gene may not be useful as genetic markers to predict the athletic performance in male Koreans.

SEQUENCE ANALYSIS AND COMPARISON OF BOVINE αS1-CASEIN GENOMIC DNA

  • Lin, C.S.;Huang, M.C.;Choo, K.B.;Tseng, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제6권4호
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    • pp.541-547
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    • 1993
  • A phage clone containing the partial ${\alpha}_{S1}$-casein gene was isolated from a bovine genomic library by using mixed probes of ovine ${\alpha}_{S1}$-, ${\beta}$- and ${\kappa}$-casein cDNAs. Restriction enzyme mapping analysis for 14.6 kb revealed that the map was in conflict with the report of Meade et al. (1990), especially in the 3'-end fragment. Sequence analysis of 12.6 kb revealed a high AT/GC ratio (1.64); we have identified eight exon sequences according to the bovine ${\alpha}_{S1}$-casein cDNA sequence. The same exon/intron splice junction sequence was observed between these exons. We suggest that the bovine ${\alpha}_{S1}$-casein gene night contain a minimum of 18 exons and the full length is approximately 18-19 kb.

MC1R gene의 PCR-RFLP를 이용한 한우.젖소고기 감별 (Analysis of Melanocortin receptor 1 (MC1R) gene differential test for beef species between Hanwoo and Holstein using polmerase chain reaction -restriction fragment length polymorphism (PCR-RFLP))

  • 서동균
    • 한국동물위생학회지
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    • 제31권3호
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    • pp.369-374
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    • 2008
  • The objective of this study was to differentiate the beef species between Hanwoo and Holstein from a total of 1,081 beef samples using PCR-RFLP of MC1R gene. When a PCR product of 403 bp specific band amplified from bovine MC1R gene sequence was digested with restriction enzyme MspA1I, Hanwoo type showed 2 bands, 220 bp and 183 bp size bands. Holstein type, however, showed three bands, 220 bp, 138 bp and 45 bp size band, respectively. The results of the differential test for beef species were as following; 7 samples (0.64%) were determined to Holstein type, of which 4 were submitted from administrative authorities, other 3 from self-collection planing, and none from civilian clients including school.

Association Study between Vitamin D Receptor Gene Polymorphism and Adult Periodontitis in Korea

  • Kang, Byung-Yong;Ha, Nam-Joo
    • Animal cells and systems
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    • 제7권2호
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    • pp.145-149
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    • 2003
  • Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RFLP) in the vitamin D receptor gene in nan periodontically healthy controls and 28 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene was detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There was no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

Polymorphism of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Native Sarabi Cows

  • Pashmi, M.;Ghorashi, S.A.;Salehi, A.R.;Moini, M.;Javanmard, A.;Qanbari, S.;Yadranji-Aghdam, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권6호
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    • pp.775-778
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    • 2006
  • Sarabi cows (n = 136) from the Sarabi Breeding Station were genotyped at bovine lymphocyte antigen (BoLA)-DRB3.2 locus by a genotyping system that used the polymerase chain reaction and restriction fragment length polymorphism. Genomic DNA was extracted from whole blood samples. A two-step polymerase chain reaction was carried out in order to amplify a 284 base-pair fragment of target gene. Nested-PCR products were digested with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Digested fragments were analyzed by polyacrylamide gel electrophoresis. Twenty-six BoLA-DRB3.2 alleles were identified with frequencies ranging from 0.4 to 15.1%. Six new allele types observed in this study have not been reported previously. Identified alleles include: BoLA-DRB3.$2^*1$, $^*2$, $^*4$, $^*6$, $^*8$, $^*12$, $^*13$, $^*14$, $^*15$, $^*16$, $^*17$, $^*23$, $^*24$, $^*25$, $^*28$, $^*32$, $^*34$, $^*35$, $^*36$, $^*37$, $^*42$, $^*46$, $^*51$, $^*kba$, $^*laa$ and $^*vaa$. Their frequencies were found to be 0.4, 0.4, 0.7, 11.4, 1.1, 1.8, 2.9, 2.2, 4.4, 9.6, 1.1, 13.6, 0.4, 0.4, 1.1, 0.7, 0.4, 6.2, 2.2, 3.7, 1.1, 7.7, 1.5, 15.1, 2.6 and 7.3% respectively. The six most frequent alleles (DRB3.2 $^*6$, $^*16$, $^*23$, $^*46$, $^*kba$ and $^*vaa$) accounted for 64.7% of the alleles in the population of this herd. Numerous studies on this locus, covering different breeds, has revealed the existence of various alleles in this locus, and new investigations have introduced novel alleles. With respect to the high number of the observed alleles in this survey and the novelty of some alleles with no previous record of reporting, it is plausible to conclude that the BoLA-DRB3.2 locus is highly polymorphic in Iranian native Sarabi cows.

Association of Chicken Growth Hormones and Insulin-like Growth Factor Gene Polymorphisms with Growth Performance and Carcass Traits in Thai Broilers

  • Nguyen, Thi Lan Anh;Kunhareang, Sajee;Duangjinda, Monchai
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권12호
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    • pp.1686-1695
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    • 2015
  • Molecular marker selection has been an acceptable tool in the acceleration of the genetic response of desired traits to improve production performance in chickens. The crossbreds from commercial parent stock (PS) broilers with four Thai synthetic breeds; Kaen Thong (KT), Khai Mook Esarn (KM), Soi Nin (SN), and Soi Pet (SP) were used to study the association among chicken growth hormones (cGH) and the insulin-like growth factor (IGF-I) genes for growth and carcass traits; for the purpose of developing a suitable terminal breeding program for Thai broilers. A total of 408 chickens of four Thai broiler lines were genotyped, using polymerase chain reaction-restriction fragment length polymorphism methods. The cGH gene was significantly associated with body weight at hatching; at 4, 6, 8, 10 weeks of age and with average daily gain (ADG); during 2 to 4, 4 to 6, 0 to 6, 0 to 8, and 0 to 10 weeks of age in $PS{\times}KM$ chickens. For $PS{\times}KT$ populations, cGH gene showed significant association with body weight at hatching, and ADG; during 8 to 10 weeks of age. The single nucleotide polymorphism variant confirmed that allele G has positive effects for body weight and ADG. Within carcass traits, cGH revealed a tentative association within the dressing percentage. For the IGF-I gene polymorphism, there were significant associations with body weight at hatching; at 2, 4, and 6 weeks of age and ADG; during 0 to 2, 4 to 6, and 0 to 6 weeks of age; in all of four Thai broiler populations. There were tentative associations of the IGF-I gene within the percentages of breast muscles and wings. Thus, cGH gene may be used as a candidate gene, to improve growth traits of Thai broilers.

아나목스 농후배양에서 암모니아 산화균의 자생 특성 (Presence and Growth of Ammonia-oxidizing Bacteria in Anaerobic Ammonium Oxidation Enrichment)

  • 배효관;;정진영
    • 한국물환경학회지
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    • 제36권3호
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    • pp.220-228
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    • 2020
  • Anaerobic ammonium oxidation (AMX) is a cost-efficient biological nitrogen removal process. The coexistence of ammonia-oxidizing bacteria (AOB) in an AMX reactor is an interesting research topic as a nitrogen-related bacterial consortium. In this study, a sequencing batch reactor for AMX (AMX-SBR) was operated with a conventional activated sludge. The AOB in an AMX bioreactor were identified and quantified using terminal restriction fragment length polymorphism (T-RFLP) and real-time qPCR. A T-RFLP assay based on the ammonia monooxygenase subunit A (amoA) gene sequences showed the presence of Nitrosomonas europaea-like AOB in the AMX-SBR. A phylogenetic tree based on the sequenced amoA gene showed that AOB were affiliated with the Nitrosomonas europaea/mobilis cluster. Throughout the enrichment period, the AOB population was stable with predominant Nitrosomonas europaea-like AOB. Two OTUs of amoA_SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments. Real-time qPCR was used to quantify AOB populations over time. Interestingly, the exponential growth of AOB populations was observed during the substrate inhibition of the AMX bacteria. The specific growth rate of AOB under anaerobic conditions was only 0.111 d-1. The growth property of Nitrosomonas europaea-like AOB may provide fundamental information about the metabolic relationship between the AMX bacteria and AOB.

Acanthamoeba pustulosa와 A. palestinensis의 동위효소 및 rDNA PCR-RFLP 양상의 유사성 (Close relatedness of Acanthomoeba pintulosa with Accnthcmoebc palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns)

  • 김영호;옥미선
    • Parasites, Hosts and Diseases
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    • 제34권4호
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    • pp.259-266
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    • 1996
  • 형태학적 제3군 가시아메바의 taxonomic validity는 아직 확실하지 않다. 이번 연구에서 제3군에 속하는 6종의 가시 아메바 즉 A. culbertsoni A. healyi A. palestinensis. A. pustulosa, A. royreba 및 A. lenticulata의 type strain들의 동위효소. 미토콘트리아 DNA 및 small subunit(ssu) rDNA의 Restriction Fragment Length Polymorphism(RFLP) 양상을 비교하여 이들의 taxonomic validity를 검토하였다. 미토콘드리아 DNA의 RFLP 양상은 분리주간에 서로 심한 차이를 보였다. A. palestinensis와 A. pustulosc는 거의 동일한 rDNA RFLP(추정 염기 치환율 2.6%) 및 동위효소의 양상을 보여 A. palestinensis와 A. pustulosc는 같은 종으로 판단되었다. 그외의 종들은 서로 아주 다양한 rDNA RFLP 및 동위효소의 양상을 나타내어 독립종으로 인정되었다.

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Duchenne Muscular Dystrophy에 관한 세포유전학 및 분자유전학적 연구 (Cytogenetic and Molecular Genetic Studies on Duchenne Muscular Dystrophy)

  • 홍해숙
    • Journal of Korean Biological Nursing Science
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    • 제7권1호
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    • pp.29-46
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    • 2005
  • Purpose ; 본 연구는 X-염색체와 관련된 장애 중에서 가장 흔하고 심한 Duchenne Muscular Dystrophy(DMD)의 세포유전학 및 분자유전학적 특성을 설명하기 위해서 DMD에 영향을 받고 있는 두 가계의 13명을 대상으로 가계도 분석과 염색체 분석 및 DNA 분석을 하였다. Method ; DNA분석은 DNA probe을 이용한 Southern blotting method로써 RFLPs와 DMD유전자 부위의 exon소실 유무를 조사하여 아래와 같은 결과를 얻었다. Conclusion ; A 염색체 분석 : 말초혈액과 양수를 표본으로 High-Resolution GTG염색에서 A가계와 B가계의 염색체 분석에서 12명의 염색체는 정상 X-염색체였으나 B가계의 I-2(DMD여성)에서 46, x,-x,+t(2:x)(q 21.1 : p21.2)로 나타난다. B. DNA분석3 : 1) RFLPs의 분석 J66,XJ-1.1,754-11로써 B가계의 RELPs(Restriction Fragment Length Polymorphisms)에서 J66/Pst I은 1.7hb(E), 1.6kb(e)을 보여 주었고 XJ-1.1/Taq I은 3.6kb(F), 3.0kb(f), 754-11/EoR I은 4.2kb(G), 2.0kb(g)의 대립인자를 나타내었다. 이상의 결과를 바탕으로 영향을 받고 있는 남자 (II-2)의 haplotype는 보인자인 어머니의 한쪽 인자를 받았으며 어머니와 딸은 보인자이고 임산부의 태아는 남아였고 태아의 인자들은 그의 할아버지로부터 물려받아 DMD에 영향을 받지 않은 것으로 진단되었다. 2) DMD 유전자의 exon 소실에 대한 분석 cDNA probe 8과 cDNA probe 2b-3으로써 소실에 대한 진단은 영향을 받은 남자(II-2)는 cDNA probe 8에서 12, 7.3, 6.6, 4.2kb에 소실이 있고 cDNA 2b-3은 1.7kb에 소실에 나타났다.

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