• Interdisciplinary Bio Central
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• 제2권4호
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• pp.14.1-14.9
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• 2010

Misidentification of cultured cell lines results in the generation of erroneous scientific data. Hence, it is very important to identify and eliminate cell lines with a different origin from that being claimed. Various methods, such as karyotyping and isozyme analysis, can be used to detect inter-species misidentification. However, these methods have proved of little value for identifying intra-species misidentification, and it will only be through the development and application of molecular biological approaches that this will become practical. Recently, the profiling of microsatellite variants has been validated as a means of detecting gene polymorphisms and has proved to be a simple and reliable method for identifying individual cell lines. Currently, the human cell lines provided by cell banks around the world are routinely authenticated by microsatellite polymorphism profiling. Unfortunately, this practice has not been widely adopted for mouse cells lines. Here we show that the profiling of microsatellite variants can be also applied to distinguish the commonly used mouse inbred strains and to determine the strain of origin of cultured cell lines. We found that approximately 4.2% of mouse cell lines have been misidentified; this is a similar rate of misidentification as detected in human cell lines. Although this approach cannot detect intra-strain misidentification, the profiling of microsatellite variants should be routinely carried out for all mouse cell lines to eliminate inter-strain misidentification.

• Journal of Animal Science and Technology
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• 제55권2호
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• pp.87-93
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• 2013

We identified four new bovine tri-nucleotide microsatellite loci and analyzed their sequence structures and genetic parameters in 105 randomly selected Korean cattle (Hanwoo). Allele numbers of the loci B17S0808, B15S6253, B8S7996, and B17S4998 were 10, 11, 12, and 29, respectively. These alleles contained a simple or compound repeat sequences with some variations. Allele distributions of all these loci were in Hardy-Weinberg equilibrium (P > 0.05). Observed heterozygosity and expected heterozygosity ranged from 0.54 (B15S6253) to 0.92 (B17S4998) and from 0.599 (B15S6253) to 0.968 (B17S4998), respectively, and two measures of heterozygosity at each locus were highly correlated. Polymorphism information content (PIC) for these 4 loci ranged from 0.551 (B15S6253) to 0.932 (B17S4998), which means that all these loci are highly informative (PIC > 0.5). Other genetic parameters, power of discrimination (PD) and probability of exclusion (PE) ranged from 0.783 (B15S6253) to 0.984 (B17S4998) and from 0.210 (B15S6253) to 0.782 (B17S4998), respectively. Their combined PD and PE values were 0.9999968 and 0.98005176, respectively. Capillary electrophoresis revealed that average peak height ratio for a stutter was 13.89% at B17S0808, 26.67% at B15S6253, 9.09% at B8S7996, and 43.75% at B17S4998. Although the degree of genetic variability of the locus B15S6253 was relatively low among these four microsatellite markers, their favorable parameters and low peak height ratios for stutters indicate that these four new tri-nucleotide microsatellite loci could be useful multiplex PCR markers for the forensic and population genetic studies in cattle including Korean native breed.

• International Journal of Industrial Entomology and Biomaterials
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• 제28권2호
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• pp.39-50
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• 2014

In order to understand the diversity and genetic relationships of silkworm strains preserved in Korea, we genotyped 78 Bombyx mori strains (Bombycidae: Lepidoptera) originating from Japan, using eight polymorphic microsatellite loci. We obtained per-locus allele numbers ranging from 5 to 16 (with an average value of 9.1), per-locus observed heterozygosity ranging from 0.13 to 1.00, and per-locus polymorphic information content ranging from 0.36 to 0.77, indicating that some loci are highly variable. Phylogenetic analysis with the eight concatenated microsatellite loci showed no clustering based on known strain characteristics and origin. Nineteen strain-specific apomorphic alleles, which discriminated 16 of the 78 silkworm strains, were obtained from eight loci. These strain-specific alleles can thus be utilized for routine discrimination of strains from Japan, without any further typing of other loci. Homozygotes were also observed at some loci (27 of 118 genotypes), which can also be used to discriminate several strains by typing a few loci. These results showed that eight microsatellite loci described herein were sufficiently variable to discriminate among the 78 silkworm strains we examined, and may be useful for future investigations of this economically important species.

• 원예과학기술지
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• 제32권6호
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• pp.853-863
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• 2014

국내외에서 재배되고 있는 딸기 100품종의 DNA profile 데이터베이스를 구축하기 위하여 다형성이 높은 microsatellite 마커의 선정과 유전적 유사도 분석을 통한 품종식별력 검정 등에 대한 연구를 수행하였다. 딸기 21품종을 274개의 microsatellite 마커로 검정하여 반복 재현성이 높은 25개의 다형성이 높은 마커를 선정하였다. 이들 마커와 국내외에서 재배되고 있는 딸기 100품종을 검정하였을 때 마커당 평균 대립유전자수는 7.50개로 나타났고, 3-13개까지 다양한 분포를 나타내었다. PIC 값은 마커의 유전자형에 따라 0.333-0.841 범위에 속하였으며 평균값도 0.706으로 높게 나타났다. Microsatellite 마커의 대립유전자를 이용하여 딸기 100 품종에 대한 계통도를 작성하였을 때 품종 육성의 계보 및 육성 지역에 따라 7개의 그룹으로 크게 나누어졌으며 2품종을 제외한 98품종이 microsatellite 마커의 유전자형에 의해 식별이 되는 것으로 나타났다. 본 연구에서 얻어진 딸기 품종별 DNA profile 데이터베이스는 품종보호 출원 품종의 재배심사 및 품종진위성과 관련된 종자분쟁을 해결하는 수단으로 유용하게 활용될 수 있을 것이다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권12호
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• pp.1696-1701
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• 2016

Japanese quail is still used as a model for poultry research because of their usefulness as laying, meat, and laboratory animals. Microsatellite markers are the most widely used molecular markers, due to their relative ease of scoring and high levels of polymorphism. The objective of the research was to determine genetic diversity and population genetic structures of selected Japanese quail lines (high body weight 1 [HBW1], HBW2, low body weight [LBW], and layer [L]) throughout 15th generations and an unselected control (C). A total of 69 individuals from five quail lines were genotyped by fifteen microsatellite markers. When analyzed profiles of the markers the observed ($H_o$) and expected ($H_e$) heterozygosity ranged from 0.04 (GUJ0027) to 0.64 (GUJ0087) and 0.21 (GUJ0027) to 0.84 (GUJ0037), respectively. Also, $H_o$ and $H_e$ were separated from 0.30 (L and LBW) to 0.33 (C and HBW2) and from 0.52 (HBW2) to 0.58 (L and LBW), respectively. The mean polymorphic information content (PIC) ranged from 0.46 (HBW2) to 0.52 (L). Approximately half of the markers were informative ($PIC{\geq}0.50$). Genetic distances were calculated from 0.09 (HBW1 and HBW2) to 0.33 (C and L). Phylogenetic dendrogram showed that the quail lines were clearly defined by the microsatellite markers used here. Bayesian model-based clustering supported the results from the phylogenetic tree. These results reflect that the set of studied markers can be used effectively to capture the magnitude of genetic variability in selected Japanese quail lines. Also, to identify markers and alleles which are specific to the divergence lines, further generations of selection are required.

• 한국어류학회지
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• 제32권2호
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• pp.39-48
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• 2020

가는돌고기(Pseudopungtungia tenuicorpa)는 8~10 cm 크기의 소형 잉어과 어류로 전 세계에서 한국의 한강 그리고 임진강에만 서식하는 멸종위기종이다. 가는돌고기는 국내 담수의 상위 포식자 중 하나인 꺽지 수컷이 돌보는 수정된 알이 있는 둥지에 탁란(brood parasitism)을 하거나 작은 바위에 생긴 틈에 산란을 하는 생식 행동을 보인다. 이 종의 특이한 생식 생태는 환경 파괴가 극심한 현대 사회에서 산란 장소를 더욱 제한할 가능성이 높아 특별한 관리와 보전 전략이 필요하다. 본 연구에서는 microsatellites와 mtDNA control region 유전자를 이용하여 가는돌고기의 종 보전 관리 전략에 필요한 개체군 수준의 유전적 다양성 등 기초자료를 확보하고자 하였다. 유전체 분석에서 얻어진 28개의 microsatellite 유전자들을 이용하여 한강의 3지역에서 채집된 67개체들의 유전자형을 밝혔다. 본 microsatellite 유전자 분석 결과, 가는돌고기는 일반적으로 알려진 담수어류의 microsatellite 다양성 정도를 훨씬 뛰어 넘는 높은 유전적 다양성을 보여주었고(평균 이형접합자 빈도 예측치=0.914; 유전자 당 평균 대립 인자 빈도=27.9), 개체군 감소나 inbreeding의 흔적은 나타나지 않았다. 그러나 북한강과 남한강 사이의 유전적 분화가 두드러졌다. 이런 유전적 구조는 14개 haplotype이 발견된 mtDNA 분석 결과에서도 유사하게 나타났다. 매우 좁은 지역에 서식하는 고유 멸종위기종에서 유전자 흐름의 제한 가능성이 나타났기 때문에, 장기적 측면에서 개체군들의 크기에 대한 고민이 필요하다. 추후 적응 유전적 분석 결과에서도 유사한 결과가 나타난다면, 북한강과 남한강 개체군들은 별도 관리가 이루어져야 하며, 복원 계획에도 이러한 유전적 구조에 대한 검토가 수반되어야 할 것이다.

• The Plant Pathology Journal
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• 제30권4호
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• pp.384-396
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• 2014

Vegetative compatibility groups (VCGs) are determined for many fungi to test for the ability of fungal isolates to undergo heterokaryon formation. In several fungal plant pathogens, isolates belonging to a VCG have been shown to share significantly higher genetic similarity than those of different VCGs. In this study we sought to examine the relationship between VCG and genetic similarity of an important cool season turfgrass pathogen, Sclerotinia homoeocarpa. Twenty-two S. homoeocarpa isolates from the Midwest and Eastern US, which were previously characterized in several studies, were all evaluated for VCG using an improved nit mutant assay. These isolates were also genotyped using 19 microsatellites developed from partial genome sequence of S. homoeocarpa. Additionally, partial sequences of mitochondrial genes cytochrome oxidase II and mitochondrial small subunit (mtSSU) rRNA, and the atp6-rns intergenic spacer, were generated for isolates from each nit mutant VCG to determine if mitochondrial haplotypes differed among VCGs. Of the 22 isolates screened, 15 were amenable to the nit mutant VCG assay and were grouped into six VCGs. The 19 microsatellites gave 57 alleles for this set. Unweighted pair group methods with arithmetic mean (UPGMA) tree of binary microsatellite data were used to produce a dendrogram of the isolate genotypes based on microsatellite alleles, which showed high genetic similarity of nit mutant VCGs. Analysis of molecular variance of microsatellite data demonstrates that the current nit mutant VCGs explain the microsatellite genotypic variation among isolates better than the previous nit mutant VCGs or the conventionally determined VCGs. Mitochondrial sequences were identical among all isolates, suggesting that this marker type may not be informative for US populations of S. homoeocarpa.

• International Journal of Industrial Entomology and Biomaterials
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• 제25권1호
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• pp.81-92
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• 2012

A total of 85 Chinese-origin silkworm strains preserved in Korea were genotyped for eight polymorphic micro-satellite loci. We obtained per-locus number of alleles, ranging from 5 to 14 with an average value of 9.5, perlocus observed heterozygosity, ranging from 0.07 to 0.99, and per-locus polymorphic information content (PIC), ranging from 0.34 to 0.82, indicating that some loci are highly variable. Phylogenetic analysis with the eight concatenated microsatellite loci showed no clustering on the basis of known strain characteristics. A total of 22 strain-specific apomorphic alleles, which discriminate 19 among 85 silkworm strains were obtained from eight loci. These strain-specific alleles, thus, can casually be utilized for the discrimination of applicable strains without any further typing of other loci. Furthermore, a substantial number of homozygote strains, represented by 27 among 76 alleles in eight loci were found. These results collectively suggest that the silkworm microsatellite DNA is actually and potentially important molecular markers for the eventual discrimination of silkworm strains that are preserved as hundreds in Korea.

• 한국유기농업학회지
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• 제12권4호
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• pp.451-461
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• 2004

In an animals, identification system has been widely used by ear tag with dummy code and blood typing for parernity. Also, genotyping methods were using for useful mean of individual identification for live animals. In the case of genotyping estimation of gene in population of korean native chicken. In this study, we tested for development of genetic markers used it possible to determination of individual identification system. The candidate genetic markers were used already bow 10 of microstalite DNA sequence information in chromosome No. 1 and 14. Result of analysis for genotyping, the number of alleles of those microstatelites DNA was shown minimal 3 to 12 and the heterozygote expression frequency range was shown from 0.617 to 0.862. In our result, effective number of allele for each microsatellites DNA was shown 3~7, and the accuracy of individual identification was shown nearly 100%, when used with 6 genetic marker. This study was about genotyping method for identification used specific genetic marker form microsatellite DNA in the brand marketing of korean native chicken. Our results suggest that genotyping method used specific genetic marker from microsatellite DNA might be very useful for determination of individual identification.

• 생명과학회지
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• 제25권1호
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• pp.16-20
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• 2015

참돔과 감성돔은 우리나라 주변에 서식하는 고유 어종으로, 참돔의 암컷과 감성돔의 수컷을 이용한 인공수정에 의하여 잡종은 생산되었으나, 자연 상태에서는 이들 간의 잡종이 보고된 바 없다. 이들 두 어종의 암수 및 타 어종을 섞어서 대형 수조에서 사육하는 과정에서 생산된 수정란을 회수하여 부화시켜 육성하는 과정에서 이들 두 종간의 잡종의 형태를 보이는 개체들이 관찰되었다. 임의로 96개체를 선택하여 두 종에 모두 적용할 수 있는 microsatellite marker를 이용하여 유전학적 분석을 실시한 결과 96개체 중 두 종의 혼합된 형태적 특징을 보이는 15개체는 참돔 암컷과 감성돔 수컷 간의 잡종으로 판명되었으며, 나머지 81개체는 감성돔 치어로 확인되었다. 사육 수조의 크기가 매우 컸으며 다른 어류들도 함께 들어 있었다는 점과, 이와 같이 유전적으로 구분되는 두 종 간의 잡종이 자연상태와 유사한 환경에서 생산되었다는 점을 고려할 때 본 연구의 결과는 자연 상태에서도 인위적인 영향이나 기후 변화에 의하여 이들의 서식지가 중복될 경우 두 종간의 잡종이 생산될 가능성이 있다는 것을 시사한다.