• 대한의생명과학회지
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• 제10권1호
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• pp.23-29
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• 2004

Nebulin is a (Mr 600∼900 kDa) large actin-binding protein specific to skeletal muscle and thought to act as a molecular template that regulates the length of thin filaments. Cardiac muscles of higher vertebrates have been shown earlier to lack nebulin. Recently, full-length nebulin mRNA transcripts have been detected in heart muscle, but at lower levels than in skeletal muscle. Nebulin expression also was detected in the kidney, eye, and otic canal, suggesting that nebulin isoforms may also be expressed in these organs. We have searched for nebulin isoforms in brain of human using PCR and Northern blot. Here, we provide evidence that nebulin mRNA transcripts are expressed in brain. Seven nebulin isoforms (B, C, D, E, F, G and H form) are obtained in human skeletal muscle and four isoforms (B, C, G and H form) in human brain cDNA library. We cloned the 1.3 kb of nebulin fragment from human adult brain library by PCR. The identity of the PCR product was confirmed by sequence analysis. The partial brain nebulin sequence was 99% identical to the skeletal muscle cDNA as determined by Blast alignment. It contains two simple-repeats HR1, HR2 and linker-repeats exon l35∼143 except exon 140. It was different from skeletal muscle B form, which contain HR1 and HR8. These data suggest that nebulin isoform diversity occurs even more extensively than previously known, likely contributing to the distinct thin filament architecture of different striated muscles.

• Molecules and Cells
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• 제25권2호
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• pp.196-204
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• 2008

Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsung-cho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to $F_2$ genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.

• Molecules and Cells
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• 제22권3호
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• pp.300-307
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• 2006

Brassica rapa ssp. pekinensis (Chinese cabbage) is an economically important crop and a model plant for studies on polyploidization and phenotypic evolution. To gain an insight into the structure of the B. rapa genome we analyzed 12,017 BAC-end sequences for the presence of transposable elements (TEs), SSRs, centromeric satellite repeats and genes, and similarity to the closely related genome of Arabidopsis thaliana. TEs were estimated to occupy 14% of the genome, with 12.3% of the genome represented by retrotransposons. It was estimated that the B. rapa genome contains 43,000 genes, 1.6 times greater than the genome of A. thaliana. A number of centromeric satellite sequences, representing variations of a 176-bp consensus sequence, were identified. This sequence has undergone rapid evolution within the B. rapa genome and has diverged among the related species of Brassicaceae. A study of SSRs demonstrated a non-random distribution with a greater abundance within predicted intergenic regions. Our results provide an initial characterization of the genome of B. rapa and provide the basis for detailed analysis through whole-genome sequencing.

• Horticulture, Environment, and Biotechnology : HEB
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• 제59권6호
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• pp.889-897
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• 2018

Currently, there is a lack of genetic markers capable of effectively detecting polymorphisms in Clematis. Therefore, we developed new markers to investigate inter- and intraspecific diversity in Clematis. Based on the complete chloroplast genome of Clematis terniflora, simple sequence repeats were explored and primer pairs were designed for all ten adequate repeat regions (cpSSRs), which were tested in 43 individuals of 11 Clematis species. In addition, the nuclear ITS region was sequenced in 11 Clematis species. Seven cpSSR loci were found to be polymorphic in the genus and serve as markers that can distinguish different species and be used in different genetic analyses, including cultivar identification to assist the breeding of new ornamental cultivars.

• 한국산림과학회지
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• 제98권5호
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• pp.568-575
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• 2009

Korean wild and forest cultivated ginseng has long been accepted as high medicinal values compared to field cultivated ginseng. Owing to the high price of Korean wild ginseng, Chinese wild and forest cultivated ginseng were smuggled and sold as Korean wild and forest cultivated ginseng. Therefore, an efficient method is required to distinguish Korean ginseng from Chinese ginseng. Microsatellites, simple sequence repeats (SSRs), are highly polymorphic loci present in DNA that consist of repeating units of base pairs. Thus SSR markers are highly advantageous for detection of small genetic variances of intra-species. In the present study, we constructed a microsatellite-enriched genomic library from South Korean wild Panax ginseng. After sequence analysis of 992 randomly picked positive colonies, 126 (12.7%) of the colonies were found to contain microsatellite sequences, and 38 primer pairs were designed. By polymorphism assessment using 36 primer pairs, 4 primers (PG409, PG450, PG491, and PG582) were shown to be polymorphic to distinguish the South Korean ginseng from the Chinese ginseng. These 4 microsatellite markers will provide powerful tools to authenticate South Korean ginseng from Chinese ginseng.

• 식물분류학회지
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• 제52권1호
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• pp.45-53
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• 2022

Halenia coreana is an endangered, endemic species that is distributed in only a few locations in Korea, such as Mts. Hwaaksan and Daeamsan. It has been recently segregated from H. corniculata, broadly distributed in cold temperate regions that include northern Japan, the Russian Far East, northeastern China, Mongolia, and eastern Europe, where population sizes are usually large. To examine the genetic diversity of H. coreana and evaluate the level of genetic differentiation of the species compared with that of H. corniculata, we surveyed 183 candidate simple sequence repeats (SSR) motif markers for H. coreana and H. corniculata from sequence data of amplified fragments of a specific length in the genome. A total of 17 genomic-SSR markers were selected to examine the levels of genetic diversity and differentiation using 17 samples of H. coreana and 60 samples of three populations of H. corniculata. The results here suggest that the genetic diversity of H. coreana is very low with a high frequency of inbreeding within its population. We found that H. coreana is genetically differentiated from H. corniculata, supporting the recognition of the geographically isolated H. coreana as a distinct species.

• 한국작물학회지
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• 제49권3호
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• pp.251-255
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• 2004

최근 비교적 재배면적이 넓은 일미벼, 대산벼, 동안벼와 이들의 모본들로 구성된 23품종의 유전적 다양성을 조사한 결과를 요약하면 다음과 같다. 1. 1999년에서 2001년에 잎도열병과 이삭도열병이 심하게 발병한 일미벼. 대산벼, 동안벼는 밀양95호를 모본으로 육성되었다. 2. 밀양95호를 모본으로 하는 품종들에 대해 SSR 마커를 이용하여 유전적 다양성을 조사한 결과 57개 유전자좌위에서 170개(평균 3.0개)의 alleles이 관찰되었으며 대립유전자수는 2개에서 7개까지 다양하였다. 특히, RM249, RM206, OSR20은 6개 이상의 대립유전자를 가져 근연의 자포니카 벼품종간의 유전적 다양성 평가에 유용하게 이용될 수 있을 것으로 판단된다. 3.증폭된 밴드의 유무를 바탕으로 품종간 유전적 거리를 산출하여 군집분석을 실시한 결과 크게 4개의 군으로 나뉘었으며 일미벼, 대산벼, 동안벼는 모본인 밀양95호와 유전적 배경이 매우 유사하였다. 4. 같은 계보상의 품종들에 대해 도열병 저항성 유전자 인근의 마커를 이용한 유전적 다양성 분석결과가 계보도와 일치하여 이들 품종들의 저항성 유전자형 또한 유사할 것으로 추청할 수 있었다. 5. 조사품종의 계보상의 allele 전이를 분석한 결과 11번 염색체의 RM254의 3번째 allele과 12번 염색체의 OSR32의 2번째 allele이 밀양95호로부터 계속 전이되었음을 알 수 있었으며 이들 alleles의 도열병 이병화와의 연관성 여부는 추후에 검토가 이루어져야할 것이다.

• 식물분류학회지
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• 제39권1호
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• pp.48-54
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• 2009

우리나라 특산식물이며 희귀식물인 만리화(물푸레나무과) 집단의 유전적 다양성을 조사하기 위하여 5 집단 84개체에 대한 ISSR (Inter-Simple Sequence Repeats) 표지자 분석을 실시하였다. 14개의 ISSR 프라이머에서 총 93개의 증폭산물을 관찰할 수 있었으며, 유전적 다양성을 나타내는 P (Percentage of polymorphic loci)값과 S.I. (Shannon's information Index)가 다른 관목류에 비해 비교적 낮게 나타났다. 집단별 유전적 다양성은 석병산집단 (P = 64.5%, S.I. = 0.281)과 설악산B집단(P = 62.4%, S.I. = 0.292)이 높았으며, 석개재집단(P = 37.6%, S.I. = 0.178)이 가장 낮았다. 전체 유전변이 중 30.6%가 집단간에 기인하는 것으로 나타났고, 나머지 69.4%는 집단내 개체간의 차이에서 기인하였다. 이러한 결과는 분포역이 매우 제한되어 있으며 불연속적으로 출연하는 희귀종이라는 점으로 미루어 볼 때 지역간의 유전자 교류가 원활하지 못해 나타난 결과라고 추정해 볼 수 있다. 유전적 거리를 이용하여 UPGMA 방법에 의한 유집분석을 실시한 결과, 집단의 지리적 격리정도와 유전적 연관성은 비교적 일치하는 경향이었다. 본 연구 결과, 만리화의 유전자원보존을 위해서는 자생지 보호와 더불어 동적인 현지외 보존(dynamic ex situ conservation)과 같은 보다 적극적인 대책이 요구되며, 더 높은 유전적 다양성을 확보하기 위해서는 소수의 집단에서 다수 개체를 선발하기보다는 집단당 소수 개체를 다수의 집단에서 선발하는 집단 위주의 보존이 더욱 효과적일 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1189-1195
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• 2014

A strain-specific identification method is required to secure Chlorella strains with useful genetic traits, such as a fast growth rate or high lipid productivity, for application in biofuels, functional foods, and pharmaceuticals. Microsatellite markers based on simple sequence repeats can be a useful tool for this purpose. Therefore, this study developed five novel microsatellite markers (mChl-001, mChl-002, mChl-005, mChl-011, and mChl-012) using specific loci along the chloroplast genome of Chlorella vulgaris. The microsatellite markers were characterized based on their allelic diversities among nine strains of C. vulgaris with the same 18S rRNA sequence similarity. Each microsatellite marker exhibited 2~5 polymorphic allele types, and their combinations allowed discrimination between seven of the C. vulgaris strains. The two remaining strains were distinguished using one specific interspace region between the mChl-001 and mChl-005 loci, which was composed of about 27 single nucleotide polymorphisms, 13~15 specific sequence sites, and (T)n repeat sites. Thus, the polymorphic combination of the five microsatellite markers and one specific locus facilitated a clear distinction of C. vulgaris at the strain level, suggesting that the proposed microsatellite marker system can be useful for the accurate identification and classification of C. vulgaris.

• 생명과학회지
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• 제14권3호
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• pp.425-428
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• 2004

ISSR마크를 사용하여 고려 인삼의 품종 및 계통간 분자적 인증과 유전적 다형현상을 조사하였다. 56개의 ISSR 프라이머 중 5개가 일곱 품종 및 계통간 명확하고 재현성이 높은 DNA분절을 나타내는 최적 프라이머로 선택되었다. 전체 43밴드는 250 bp - 1,700 bp의 분자량을 가지며 프라이머당 8.6개의 밴드를 나타내었다. 고려 인삼에서 다형현상 정도는 20.9%였다. 특히 천풍 품종이 가장 높은 다형현상을 나타낸 반면 다른 품종은 거의 다형현상을 나타내지 않았다. 결론적으로 DNA수준에서 ISSR마크로 천풍이 다른 고려 인삼의 품종 및 계통인 연풍, 황숙종, 자경종과 구분에 이용될 수 있음이 판명되었다.