가시나무류의 RAPD 유전분석을 위한 Genomic DNA 최적 추출조건 및 RAPD Primer 선별
Optimal Genomic DNA Extraction Conditions and Primer Selection for RAPD Analysis of Evergreen Quercus spp.
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유전분석을 위해서, CTAB buffer를 이용하여 가시나무류 4종의 genomic DNA를 분리하였다. CTABbuffer를 이용하여 분리한 genomic DNA의 순도는 Edward buffer를 이용했을 때보다 더 높게 나타났다. 가시나무(Q. myrsinaefolia)로부터적정 순도 gDNA는 2% CTAB에 0, 1 또는 2% PVP가 첨가된 buffer를이용했을 때 얻어졌다. 종가시나무(Q. glauca)로부터 1% CTAB buffer와 1% PVP를 첨가한 buffer를 이용하여 얻은 gDNA 순도는 1.84±0.02(A260/280)였다. 참가시나무(Q. salicina)의 적정순도 gDNA는 2%CTAB와 1 또는 5% PVP가 첨가된 buffer를 이용하여 분리할 수 있었다. 2% CTAB와 2% PVP가 첨가된buffer를 이용했을 때, 졸가시나무(Q. phillyraeoides)의 gDNA의 순도는 1.85±0.01(A260/280)였다. 18개의 RAPD primer를 사용하여 PCR을 수행하였을 때, 가시나무에서는 15개 primer에 의해 53개의 다형질DNA가 발견되었다. 종가시나무에서는 11개의 primer에 의해 40개의 다형질 DNA가 나타났다. 참가시나무의 경우 16개의 primer에 의해 50개의 증폭된 DNA밴드를 확인 할 수 있었다. 졸가시나무에서 14의primer가 증폭반응을 일으켰고, 53개의 다형질 DNA가 나타났다. 이 결과들은 가시나무류의 유전분석에이용할 수 있다.
Abstract
For genetic analysis of four species of evergreen Quercus spp, their genomic DNA were isolatedusing CTAB buffer method. The purity values of genomic DNA of each species isolated usingCTAB buffer were higher than those isolated using Edward buffer. The adequate purity of genomicDNA of Quercus myrsinaefolia was obtained using the buffer containing 2% CTAB and 0, 1 or2% PVP. The purity of gnomic DNA of Q. glauca was 1.84±0.02(A260/280) when the buffercontaining 1% CTAB and 1% PVP was used. Adequate genomic DNA purity of Q. salicina couldbe isolated using the buffer containing 2% CTAB and 1 or 5% PVP. The purity of genomic DNAof Q. phillyraeoides was 1.85±0.01(A260/280), and it was obtained when the buffer 2% CTAB and2% PVP was used. When the PCR reactions using 18 RAPD primers were performed, 15 primerproduced 53 amplified fragments of Q. myrsinaefolia. Eleven primers produced 40 polymorphicDNA of Q. glauca. For amplification of random polymorphic DNA of Q. salicina, 16 primersproduced 50 bands. Of Q. phillyraeoides, 53 polymorphic bands were amplified by 14 randomprimers. These results can be used for genetic analysis of evergreen Quercus spp.
참고문헌 (23)
- Aykut Y, "Molecular diversity among Turkish oaks(Quercus) using random amplified polymorphic DNA(RAPD) analysis", African J. of Biotechnology, 12, pp.6358~6365 (2013)[DOI]
- Bacilieri R, "Comparison of morphological characters and molecular markers for the analysis of hybridization in sessile and pedunculate oak", Ann. Sci. For., 53, pp.79~91 (1996)[DOI]
- Edwards K, "A simple and rapid method for the preparation of plant genomic DNA for PCR analysis", Nucl. Acid Res., 19, pp.1349~ (1991)[DOI]
- Fraga J, "Optimization of random amplified polymorphic DNA techniques for its use in genetic studies of Trichomonasvaginalis isolates", Infect. Genet. Evol., 2, pp.73~75 (2002)[DOI]
- Gonzalez-rodriguez A, "Morphological and rapd analysis of hybridization between Quercus affinis and Q. laurina(Fagaceae), two mexican red oaks", American J. of Botany, 91, pp.401~409 (2004)[DOI]
- 구현아, "참가시나무 추출물의 항균 활성 및 크림 안정성 평가", 한국미생물·생명공학회지, 42(2), pp.145~151 (2014), 한국미생물·생명공학회[DOI]
- Henry R, "Plant Genotyping: the DNA Fingerprinting of Plants", pp.239~249 (2001), CAB International
- Kim JJ, "Comparison of phenolic compounds content in indeciduous Quercus species", J. of Medicinal Plants Research, 6, pp.5228~5239 (2012)[DOI]
- "Forest Resources"[URL]
- 이혜진, "참가시나무의 항산화 효과 및 활성 성분 분석", 대한화장품학회지, 37(2), pp.143~152 (2011), 사단법인 대한화장품학회
- McGuigan PJ, "Propagation of Quercus myrsinaefolia and Quercus canbyi by stem cuttings", J. Environ. Hort., 14, pp.217~220 (1996)
- Michiels A, "Extraction of high-quality genomic DNA from latex-containing plants", Analytical Biochemistry, 315, pp.85~89 (2003)[DOI]
- Pandey A, "High-molecular-weight DNA extraction from six Quercus species of Kumaun Himalaya India", International J. of Advanced Research, 3, pp.30~34 (2015)
- Petit RJ, "Fagaceae trees as models to integrate ecology, evolution and genomics", New Phytologist, 197, pp.369~371 (2013)[DOI]
- Porebski S, "Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components plant", Molecular Biology Reporter, 15, pp.8~15 (1997)[DOI]
- Sahu SK, "DNA Extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol", ISRN Molecular Biology, 2012, pp.205049~ (2012)
- Sharma P, "An improved method of DNA isolation from polysaccharide rich leaves of Boswellia serrata Roxb", Indian J. of Biotechnology, 11, pp.67~71 (2012)
- Shin HC, "Warm temperate tree species in Korea", pp.218~ (2006)
- Song JL, "Protective effects of Quercus salicina on alloxan-induced oxidative stress in HIT-T15 pancreatic <TEX>${\beta}$<", TEX> cells, 2013, pp.5, 947~951
- 신수정, "졸가시나무 탄화물 분석", 목재공학, 41(3), pp.181~186 (2013), 한국목재공학회[DOI]
- Souza HAV, "Isolation of high quality and polysaccharide-free DNA from leaves of Dimorphandra mollis (Leguminosae), a tree from the Brazilian Cerrado", Genetics and Molecular Research, 11, pp.756~764 (2012)[DOI]
- Tuyen PT, "Antioxidant capacity and phenolic contents of three Quercus species", International Letters of Natural Sciences, 54, pp.85~99 (2016)[DOI]
- Yakovlev IA, "Genetic differentiation of pedunculate oak Quercus robur L. in the european part of Russia based on RAPD markers", Russian J. of Genetics, 38, pp.148~155 (2002)[DOI]