Species and Sex Identification of the Korean Goral (Nemorhaedus caudatus) by Molecular Analysis of Non-invasive Samples

김백준,이윤선,안정화,박한찬,Hideo Okumura,이항,민미숙 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.3

Korean long-tailed goral (Nemorhaedus caudatus) is one of the most endangered species in South Korea. However, detailed species distribution and sex ratio data on the elusive goral are still lacking due to difficulty of identification of the species and sex in the field. The primary aim of this study was to develop an economical PCR-RFLP method to identify species using invasive or non-invasive samples from five Korean ungulates: goral (N. caudatus), roe deer (Capreolus pygargus), feral goat (Capra hircus), water deer (Hydropotes inermis) and musk deer (Moschus moschiferus). The secondary aim was to find more efficient molecular sexing techniques that may be applied to invasive or non-invasive samples of ungulate species. We successfully utilized PCR-RFLP of partial mitochondrial cytochrome b gene (376 bp) for species identification, and sex-specific amplification of ZFX/Y and AMELX/Y genes for sexing. Three species (goral, goat and water deer) showed distinctive band patterns by using three restriction enzymes (XbaI, StuI or SspI). Three different sexing primer sets (LGL331/335 for ZFX/Y gene; SE47/48 or SE47/53 for AMELX/Y gene) produced sex-specific band patterns in goral, goat and roe deer. Our results suggest that the molecular analyses of non-invasive samples might provide us with potential tools for the further genetic and ecological study of Korean goral and related species.

Sex Identification in Cinereous Vulture (Aegypius monachus) from Feather and Blood Samples: A Case Report

석성훈,강선영,한재익,임영빈,유한상,연성찬 한국임상수의학회 2020 한국임상수의학회지 Vol.37 No.1

Twenty-four cinereous vultures that had been taken to a wildlife center due to starvation and exhaustion were studied to evaluate approaches for determining sex. Coelioscopy was performed to identify sexes of two vultures, whereas, DNA testing was performed to identify the sexes of the 24 vultures. Testes and ovaries could be unambiguously identified with an endoscope and DNA analyses could identify sex sex in most, but not all of the specimens. Although the coelioscopy examination can unambiguously confirm sex, the approach is invasive and requires anesthesia. Thus, coelioscopic examination should only be performed when sex cannot be determined through DNA analysis.

웅성 특이적 SRY 및 ZFY 유전자를 이용한 쇠고기 성(性) 판별

신성철,정구용,정의룡,Shin, Sung-Chul,Chung, Ku-Young,Chung, Eui-Ryong 한국축산식품학회 2007 한국축산식품학회지 Vol.27 No.3

본 연구는 포유류의 Y-염색체상에 존재하는 SRY 및 ZFY의 웅성 특이적 성 결정 유전자를 이용하는 PCR 기법으로 쇠고기 성판별 기술을 개발하고자 수행하였다. 성 결정 유전자 영역의 특정 염기서열을 포함하는 primer를 각각 설계 합성하고 이들 primer를 이용하여 PCR증폭을 실시한 다음, 각각의 증폭산물을 1.5% agarose gel에 전기영동하여 웅성 특이적 DNA band의 증폭여부를 확인하였다. SRY 유전자에서 웅성개체 쇠고기는 1,348 bp 크기의 단편을 가진 DNA band가 검출되었으나, 자성개체의 경우 DNA band가 전혀 검출되지 않은 것을 확인 할 수 있었다. 또한, ZFY 유전자에서 웅성개체의 쇠고기는 979 bp 크기의 단편을 가진 DNA band가 모두 검출되었으나, 자성개체의 쇠고기에서는 역시 DNA band가 전혀 검출되지 않았다. 즉, SRY 및 ZFY 유전자는 모두 숫소에서 유래한 쇠고기에서 웅성 특이적인 DNA band가 정확히 검출된 반면 암소에서 유래한 쇠고기에서는 웅성 특이적 DNA band가 전혀 검출되지 않았다. 또한 쇠고기 성 감별법을 도축 후 가공 유통판매단계에 실제 활용 가능성을 검증하고자 시중에 유통되고 있는 등심 및 갈비 포장육 350시료를 무작위 추출하여 성 판별을 실시한 결과 숫소가 252시료(72%) 그리고 암소가 98두(28%)로 조사되었다. 따라서 본 연구에서 개발한 SRY 또는 ZFY의 웅성 특이적 성 결정 유전자를 이용하는 쇠고기 성 감별기술은 생산단계는 물론 도축 후 가공 유통 판매되고 있는 모든 쇠고기의 암수 성감별을 위한 유용한 DNA 표지인자로 활용할 수 있을 것으로 기대된다. The objective of this study was to develop a rapid and reliable method for the sex determination of beef using the PCR(polymerase chain reaction) technique. We have used two bovine sex determining genes, SRY and ZFY, on the Y-chromosome to identify the sex of Hanwoo and Holstein beet. We attempted to amplify 1,348 bp and 979 bp fragments from male and female genomic DNA corresponding to the SRY and ZFY genes, respectively, using male specific primers. The amplified PCR products were separated by electrophoresis in a 1.5% agarose gel to detect a male specific DNA band. When DNA from male beef was amplified with primers specific for the SRY gene, a DNA band of 1,348 bp was present in all of the male samples, but absent from all of the female samples. Also, when DNA from male beef was amplified with primers specific for the ZFY gene, a DNA band of 979 bp was observed in all of the male samples, but absent from all female samples. In conclusion, the bovine SRY and ZFY genes are typically found only in male beef. For the practical application of this method for the sexing of commercial beef at the processing and marketing stages after slaughter. a total of 350 beef samples collected randomly from local markets were analyzed for sex determination. The proportions of male and female samples were 252 (72%) and 98 (28%), respectively. Therefore. the SRY and ZFY genes. which are specific for the Y-chromosome, may be useful sex-diagnostic DNA markers to distinguish male meat from female meat.

멸종위기종 두루미( Grus japonensis) 유조의 성별확인

이상직(Sang-Jik Lee),박소영(So-young Park),박희천(Hee-Cheon Park) 한국조류학회II 2010 한국조류학회지 Vol.17 No.4

서식지외보전기관인 조류생태환경연구소에서 멸종위기종 두루미(Grus japonensis)의 복원을 위해 2010년에 인공부화한 유조 4개체의 성별을 확인하기 위하여, crane sex-linked DNA at W chromosome (CSL-W)를 primer set (mCSL01/mCSL02)를 사용하여 약 420bp의 CSL-W fragment를 증폭한 결과, RCC1, RCC4는 암컷, RCC2, RCC3는 수컷으로 확인되었다. For the restoration of Red-crowned Crane (Grus japonensis), endangered species, at the Institute of Ornithology designated by the Ministry of Environment as the ex-situ conservation organization, we had analyzed the sex identification of four chicks of Red-crowned Crane incubated in 2010. By the results of amplification using primer set (mCSL01/mCSL02) on the crane sex-linked DNA at W chromosome (CSL-W), RCC1 and RCC4 were female, whereas RCC2 and RCC3 were male.

Sex Identification of the First Incubated Chicks of the Crested Ibis Nipponia nippon in Korea

Kyung A Kim(김경아),Jae Seok Cha(차재석),Tae Jwa Kim(김태좌),Kyung Min Kim(김경민),Hee Cheon Park(박희천) 한국생명과학회 2011 생명과학회지 Vol.21 No.5

세계적 멸종위기종인 따오기(Nipponia nippon)는 2008년 10월에 중국에서 1쌍이 도입된 후 한국최초로 인공번식에 성공하였다. 본 연구는 따오기의 sex-related gene과 Chromodomain Helicase DNA Binding Protein gene(CHD gene)을 가지고 polymerase chain reaction (PCR)을 수행하여 새로 태어난 따오기 유조의 성별을 확인하고자 하였다. 본 연구에서는 따오기의 성별 확인을 위해 PCR후 제한효소의 처리 방법과 P2과 P8를 이용한 PCR 방법을 실시하였을 때 더 정확한 결과가 나타남을 알 수 있었다. 그리고 CHD gene의 염기서열을 선행연구와 비교해 본 결과, 암컷의 염기서열에서 1~2 base pairs 차이가 나타남을 알 수 있었다. In October 2008, a pair of Crested ibis Nipponia nippon, an endangered avian species in the world, was donated to Korea from China. They have since been the subject of a successful program to incubate chicks for the first time in South Korea. This study was carried out to determine the sex of chicks from the Crested ibis through polymerase chain reaction (PCR) using the sex-related gene and the chromodomain helicase DNA binding protein (CHD) gene. The result of the CHD gene, which was used with a single set of primers and a restriction enzyme treatment after the PCR process, was more accurate in identifying the gender of the Crested ibis. In addition, we compared the CHD gene sequences with the previously reported sequences and found 1~2 different bases between females (CI2, CI4, CI5, and CI6) than in studies previously reporting female sequences.

제한효소 SacⅡ의 Isoschizomer인 SexⅢ를 생성하는 토양 Streptomyces 균주의 수리 동정

배무,서원나,박정옥,김형태,이계준 이화여자대학교 생명과학연구소 1993 생명과학연구논문집 Vol.4 No.-

제한효소인 SexⅢ을 생성하는 Streptomyces속의 한 균주(Streptomyces sp.58)를 토양으로부터 분리하여 형태적 관찰 및 수리동정을 실시하였다. 50여개의 분류 단위 형질을 분석하였고 이 실험 결과를 TAXON program에 적용하여 종의 수리 동정을 실시하였다. 그 결과 분리균은 Streptomyces의 제5주군집에 속하는 것으로 나타나 Streptomyces exfoliatus의 한 균주로 동정하였다. Numerical identification was carried out for an isolate of Streptomyces sp. 58 producing a new restriction endonuclease SexⅢ. Fifty taxonomic unit characters were tested and the data were analyzed numerically using the TAXON program. The isolate was identified to the major 5 or Streptomyces. Therefore, it was concluded that the isolate was identified to be a member of Streptomyces exfoliatus.

Determination of sex from various hand dimensions of Koreans

Jee, Soo-Chan,Bahn, Sangwoo,Yun, Myung Hwan Elsevier 2015 Forensic science international Vol.257 No.-

<P><B>Abstract</B></P> <P>In the case of disasters or crime scenes, forensic anthropometric methods have been utilized as a reliable way to quickly confirm the identification of victims using only a few parts of the body. A total of 321 measurement data (from 167 males and 154 females) were analyzed to investigate the suitability of detailed hand dimensions as discriminators of sex. A total of 29 variables including length, breadth, thickness, and circumference of fingers, palm, and wrist were measured. The obtained data were analyzed using descriptive statistics and <I>t</I>-test. The accuracy of sex indication from the hand dimensions data was found using discriminant analysis. The age effect and interaction effect according to age and sex on hand dimensions were analyzed by ANOVA. The prediction accuracy on a wide age range was also compared. According to the results, the maximum hand circumference showed the highest accuracy of 88.6% for predicting sex for males and 89.6% for females. Although the breadth, circumference, and thickness of hand parts generally showed higher accuracy than the lengths of hand parts in predicting the sex of the participant, the breadth and circumference of some finger joints showed a significant difference according to age and gender. Thus, the dimensions of hand parts which are not affected by age or gender, such as hand length, palm length, hand breadth, and maximum hand thickness, are recommended to be used first in sex determination for a wide age range group. The results suggest that the detailed hand dimensions can also be used to identify sex for better accuracy; however, the aging effects need to be considered in estimating aged suspects.</P>

Developing DNA Markers based on Male-Specific Chromosomal Regions for Selecting Male Plants in Hop (Humulus lupulus)

하태현,유재일,이소영,강시용 한국육종학회 2024 Plant Breeding and Biotechnology Vol.12 No.-

Hops are dioecious, with only female plants cultivated for resin production and primarily used in brewing. Therefore, early sex identification at the seedling stage is crucial for breeding and cultivation. Molecular marker-assisted selection facilitates rapid and reliable sex identification of hops. We developed molecular male markers by leveraging sequence information from male-specific regions in a designated public database. To assess the accuracy of sex identification using the newly generated markers, we performed PCR analysis on four cultivars of hops with known sexes. Seventy-eight percent of the tested PCR primers correlated with the male sex phenotype. Following optimization, four primer pairs were successfully converted into male-specific PCR markers.

Development of Species Identification Method of Trapped and Damaged Sample using Direct PCR

Kim Ju Il,Kwon Min,Kim Jeom Soon,Lee Yong Gyu,Shim Jae Dong,Kim Seong Hee 한국응용곤충학회 2012 한국응용곤충학회 학술대회논문집 Vol.2012 No.10

Lepidopteran pests monitoring in adult stage was generally performed using delta or corn typed trap including rubber septa impregnated sex pheromone (lure). Sometimes, unfortunately trapped samples were severly damaged because of biotic and/or abiotic environments such as micro-organism, predator and rain, sticky material, respectively. In our case, we monitored potato tuber moth, PTM, Phthorimaea operculella distribution during 2009~2012 in Korea. However, we encountered unexpected problem, another species can be trapped in species specific sex pheromone trap. Therefore, species confirmation was needed in trapped samples. Here we developed confirmation method by direct PCR (without DNA extraction) or sequencing methods which trapped samples that cannot identified by morphologically. We designed multi-plex PCR universal primers and species specific primers in rRNA region because to check the success of PCR and species identification. This direct PCR method can be applied in other species confirmation which monitored using pheromone trap.

Molecular identification and sex determination for unidentified specimens of artiodactyl mammals collected in Mt. Yongmasan, Seoul, South Korea

Ju-Young Moon,Chea-Un Cho,Tae-Wook Kim,Jang-ick Son,Gyu-cheol Kim,Yong-hak Lee,Seung-Jun Jeong,Dong-ju Song,Sang-Hyun Han 강원대학교 산림과학연구소 2018 강원대학교 산림과학연구소 학술대회 Vol.2018 No.09

Recently several observations have been reported on the mass-media about habitation of long-tailed goral (Naemorhedus caudatus) in eastern borderline (Mt. Yongmasan) of the metropolitan city, Seoul. Non-invasive and unidentified specimens (hairs and pellets) were collected and we tested molecular analyses such as species identification, molecular sexing and individual counting. Nucleotide sequences of mitochondrial DNA cytochrome B and cytochrome oxidase I genes showed that those samples were water deer (Hydropotes inermis) and long-tailed goral. In addition from the results of molecular sexing we found a male and a female long-tailed goral DNA samples using male-specific sex-related region Y gene and sexually heterogenous zinc finger -X and -Y genes, respectively. The genotypes of microsatellite markers differentiated the DNA samples into a water deer and two individuals of long-tailed goral. Combining whole molecular information, we finalized that those specimens might be from at leat a water deer and two long-tailed goral. From the camera trapping (CT) survey, we also took some photos of male goral supposed as the same individual from several locations. However, we could not caught the female one at that time by CT. Although we have no direct evidence on habitation of female goral until now, our molecular assay results indicated that both male and female goral inhabit Mt. Yongmasan, Seoul. Anyway, this is the new and first record about habitation of long-tailed goral nearby the metropolitan city Seoul where might be the farthest away to the west from the main habitat (far eastern mountainous ranges of South Korea). Our findings concluded that further survey should be carry out to estimate the number of individuals by molecular test and camera trapping together, and it is necessary to establish a conservation program for the long-tailed goral in Mt. Yongmasan, Seoul.