• 한국발생생물학회지:발생과생식
• /
• 제24권4호
• /
• pp.327-335
• /
• 2020

Genomic DNA (gDNA) extracted from two populations of natural and cultured river pufferfish (Takifugu obscurus) was amplified by polymerase chain reaction (PCR). The complexity of the fragments derived from the two locations varied dramatically. The genetic distances (GDs) between individuals numbered 15 and 12 in the cultured population was 0.053, which was the lowest acknowledged. The oligonucleotide primer OPC-11 identified 88 unique loci shared within each population reflecting the natural population. The OPC-05 primer identified 44 loci shared by the two populations. The average band-sharing (BS) values of individuals in the natural population (0.683±0.014) were lower than in those derived from the cultured population (0.759±0.009) (p<0.05). The shortest GD demonstrating a significant molecular difference was found between the cultured individuals # 15 and # 12 (GD=0.053). Individual # 02 of the natural population was most distantly related to cultured individual # 22 (GD=0.827). A cluster tree was built using the unweighted pair group method with arithmetic mean (UPGMA) Euclidean GD analysis based on a total of 578 various fragments derived from five primers in the two populations. Obvious markers identified in this study represent the genetic structure, species security, and proliferation of river pufferfish in the rivers of the Korean peninsula.

• Clinical and Experimental Reproductive Medicine
• /
• 제49권4호
• /
• pp.225-238
• /
• 2022

The ultimate goal of human assisted reproductive technology is to achieve a healthy pregnancy and birth, ideally from the selection and transfer of a single competent embryo. Recently, techniques for efficiently evaluating the state and quality of preimplantation embryos using time-lapse imaging systems have been applied. Artificial intelligence programs based on deep learning technology and big data analysis of time-lapse monitoring system during in vitro culture of preimplantation embryos have also been rapidly developed. In addition, several molecular markers of the secretome have been successfully analyzed in spent embryo culture media, which could easily be obtained during in vitro embryo culture. It is also possible to analyze small amounts of cell-free nucleic acids, mitochondrial nucleic acids, miRNA, and long non-coding RNA derived from embryos using real-time polymerase chain reaction (PCR) or digital PCR, as well as next-generation sequencing. Various efforts are being made to use non-invasive evaluation of embryo quality (NiEEQ) to select the embryo with the best developmental competence. However, each NiEEQ method has some limitations that should be evaluated case by case. Therefore, an integrated analysis strategy fusing several NiEEQ methods should be urgently developed and confirmed by proper clinical trials.

• Journal of Animal Science and Technology
• /
• 제58권11호
• /
• pp.40.1-40.5
• /
• 2016

Background: Currently about 26,000 horses are breeding in Korea and 57.2% (14,776 horses) of them are breeding in Jeju island. According to the statistics published in 2010, the horses breeding in Jeju island are subdivided into Jeju horse (6.1%), Thoroughbred (18.8%) and Halla horse (75.1%). Halla horses are defined as a crossbreed between Jeju and Thoroughbred horses and are used for horse racing, horse riding and horse meat production. However, little research has been conducted on Halla horses because of the perception of crossbreed and people's weighted interest toward Jeju horses. Method: Using 17 Microsatellite (MS) Markers recommended by International Society for Animal Genetics (ISAG), genomic DNAs were extracted from the hair roots of 3,880 Halla horses breeding in Korea and genetic diversity was identified by genotyping after PCR was performed. Results and conclusion: In average, 10.41 alleles (from 6 alleles in HTG7 to 17 alleles in ASB17) were identified after the analysis using 17 MS Markers. The mean value of $H_{obs}$ was 0.749 with a range from 0.612(HMS1) to 0. 857(ASB2). Also, it was found that $H_{\exp}$ and PIC values were lowest in HMS1 (0.607 and 0.548, respectively), and highest in LEX3(0.859 and 0.843, respectively), and the mean value of $H_{\exp}$ was 0.760 and that of PIC was 0.728. 17 MS markers used in this studies were considered as appropriate markers for the polymorphism analysis of Halla horses. The frequency for the appearance of identical individuals was $5.90{\times}10^{-20}$ when assumed as random mating population and when assumed as half-sib and full-sib population, frequencies were $4.08{\times}10^{-15}$ and $3.56{\times}10^{-8}$, respectively. Based on these results, the 17 MS markers can be used adequately for the Individual Identification and Parentage Verification of Halla horses. Remarkably, allele M and Q of ASB23 marker, G of HMS2 marker, H and L of HTG6 marker, L of HTG7 marker, E of LEX3 marker were the specific alleles unique to Halla horses.

• Molecules and Cells
• /
• 제26권3호
• /
• pp.250-257
• /
• 2008

Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.

• 한국자원식물학회지
• /
• 제17권3호
• /
• pp.265-271
• /
• 2004

AFLP marker의 유용성 을 알아보고자 재배콩과 야생콩을 대상으로 유전적 다양성과 유전분리 현상을 분석하였다. 공시 재료들의 polymorphism은 재배 콩과 야생 콩에서 각각 평 균 2 9%와 12.2%의 polymorphism을 보였으며, 재배 콩과 야생 콩에서 공히 유전적 다양성을 보인 DNA단편은 11개 primer 평균 24개를 나타내었다. Primer 조합별로도 polymorphism에 다양한 차이가 있었는데 평균 22.9%로 13.0-38.5%의 변이를 나타내었다. 재배 콩 간의 교잡후대(화엄풋콩 ${\times}$ PI417479) F$_2$집단에서 AFLP marker의 유전분리 양상을 분석한 결과 3 : 1의 분리 비를 따르는 것으로 판단되었다.

• 한국육종학회지
• /
• 제43권5호
• /
• pp.360-367
• /
• 2011

Beta-carotene producing transformants were produced in the background of 'Nagdongbyeo', a Japonica rice cultivar. Introgression of the carotenoid locus in the transformant, PAC4-2 into the elite cultivar 'Ilpumbyeo' was started. To initiate a backcrossing program, we surveyed 220 SSR markers and found that 38% of them were polymorphic between 'Ilpumbyeo' as a recurrent parent and the PAC4-2 as a recipient parent. The selection strategy comprising foreground and background selection was employed. First, foreground selection was practiced in $BC_1$, $BC_2$, and $BC_3$ generations using the transgene specific PCR-based marker in addition to visual scoring of the seed color. Marker-based background selection combined with phenotypic selection was employed from $BC_3F_2$ to $BC_3F_4$ generations. Blast search indicated that the transgene PAC4-2 was located between SSR markers, RM6 and RM482. 240 $BC_3F_3$ and 63 $BC_3F_4$ lines were evaluated for four agronomic traits including days to heading. Most of the lines were similar to Ilpumbyeo in agronomic traits evaluated. The percentage of PAC4-2 genome ranged from 4% to 21% with a mean of 12.5%, which was higher than the expected for an unselected $BC_3$ backcross population. This could be explained by the fact that two genes for beta-carotene and the stripe virus resistance were targeted in this study. We selected 10 representative $BC_3F_5$ lines from 63 $BC_3F_4$ lines based on agronomic traits and carotenoids content. The selection strategy would be appropriate for the introgression of beta-carotene gene in a breeding program.

• Genomics & Informatics
• /
• 제15권1호
• /
• pp.2-10
• /
• 2017

Early detection and proper management of kidney rejection are crucial for the long-term health of a transplant recipient. Recipients are normally monitored by serum creatinine measurement and sometimes with graft biopsies. Donor-derived cell-free deoxyribonucleic acid (cfDNA) in the recipient's plasma and/or urine may be a better indicator of acute rejection. We evaluated digital PCR (dPCR) as a system for monitoring graft status using single nucleotide polymorphism (SNP)-based detection of donor DNA in plasma or urine. We compared the detection abilities of the QX200, RainDrop, and QuantStudio 3D dPCR systems. The QX200 was the most accurate and sensitive. Plasma and/or urine samples were isolated from 34 kidney recipients at multiple time points after transplantation, and analyzed by dPCR using the QX200. We found that donor DNA was almost undetectable in plasma DNA samples, whereas a high percentage of donor DNA was measured in urine DNA samples, indicating that urine is a good source of cfDNA for patient monitoring. We found that at least 24% of the highly polymorphic SNPs used to identify individuals could also identify donor cfDNA in transplant patient samples. Our results further showed that autosomal, sex-specific, and mitochondrial SNPs were suitable markers for identifying donor cfDNA. Finally, we found that donor-derived cfDNA measurement by dPCR was not sufficient to predict a patient's clinical condition. Our results indicate that donor-derived cfDNA is not an accurate predictor of kidney status in kidney transplant patients.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2003년도 춘계 학술대회지
• /
• pp.42-42
• /
• 2003

• 생명과학회지
• /
• 제30권10호
• /
• pp.912-918
• /
• 2020

본 연구는 재래흑염소와 교잡종 염소 총 304두를 대상으로 Microsatellite (MS) marker의 대립유전자형 분석을 통해 염소의 개체식별과 친자확인을 목적으로 실시하였다. 각 MS marker 별 대립유전자형의 다형성을 토대로 11종의 MS marker를 선발하였다. 선발된 MS marker를 사용할 경우 동일한 유전자형을 가진 개체가 출현할 확률이 무작위, 반형매 교배집단에서 각각 5.58×10^-10, 1.15×10^-7으로 분석되었다. 또한 친자감정 확률은 부모의 정보가 있을 경우 0.999996, 부모의 정보가 없을 경우 0.999833으로 분석되어 국내에서 사육하고 있는 염소들의 개체식별 및 친자확인이 가능할 것으로 사료된다. 또한 국내 재래흑염소 4 계통과 교잡종 염소들 간의 혈연관계 분석을 통해 국내 재래흑염소의 유전적 특성을 확인하였다. 본 연구의 결과는 염소의 개량 기반 구축에 필요한 개체관리와 친자감별 및 향후 염소고기의 생산 이력 구축에 유용하게 활용 할 수 있을 것으로 판단된다.

• 원예과학기술지
• /
• 제28권4호
• /
• pp.627-637
• /
• 2010

최근 들어 우리나라에서 토마토 소비가 급증하고 있으며 많은 토마토 품종이 시장에서 거래되고 있다. 그러나 토마토 품종 육성에 이용되는 부모 계통의 유전적 다양성이 낮아 형태적인 특성에 의한 토마토 품종의 구분은 매우 어려운 현실이다. 이에 따라 토마토의 품종을 구별해 낼 수 있는 분자표지의 개발이 필요한 실정이다. 본 연구에서는 SNP를 탐색하고 토마토 품종 구분을 위한 SNP 마커를 개발하였다. SNP분자표지는 고추 유전체 서열로부터 파생된 COS II 분자표지와 인트론 기반 분자표지를 기반으로 선발되었으며, HRM분석을 통해 다형성을 테스트 하였다. 전체 628개의 프라이머 조합 가운데 PCR을 통해 크기가 500bp 이하의 단일 밴드가 증폭된 417개의 프라이머 조합을 선발하였다. 417개의 프라이머 조합을 이용해 4개의 토마토 계통을 대상으로 HRM 분석을 실시하였으며, 다형성을 보인 70개의 프라이머 조합을 선발하였다. 70개의 프라이머 조합을 이용하여 32개의 토마토 품종을 대상으로 HRM 분석을 실시하였다. HRM분석을 통해 총 11개의 SNP 분자표지가 선발되었으며, 이 분자표지를 이용해 시판중인 32개의 토마토 품종을 모두 구분할 수 있었다.