The long-tailed goral (Naemorhedus caudatus) is an endangered animal species in all its habitats worldwide, including South Korea. The imbalanced sex ratio in fragmented habitats is closely associated with extinction. Therefore, sex identification using wild animal samples would be necessary. However, only a few studies have been reported about the sex identification of gorals. In this study, we thus aimed at comparing the efficiency of sex identification using various goral sample types as templates and the amelogenin (AMEL) and DEAD-box polypeptide 3 (DDX3) genes as target sequences. We extracted DNA from goral feces, tissues, and blood samples, then amplified the AMEL (SE47/SE48 and SE47/SE53 primer pairs) and DDX3 genes for sex identification, comparing the goral DDX3X and DDX3Y target sequences to those in cattle. Our results indicated that the tissue- and blood sample-derived AMEL amplicons showed an unspecific band pattern containing the sex-specific band in the case of both primer pairs we used, whereas the DDX3 amplicon showed only the sex-specific band. In the case of the feces samples, only the sex-specific band was amplified using both the AMEL and DDX3 primer pairs. However, we found that the DDX3 amplicon exhibited a clearer band pattern than the AMEL amplicon. Then, we compared the DDX3X and DDX3Y target sequences between cattle and gorals. We found 5 and 8 nucleotide differences in the DDX3X and DDX3Y sequences, respectively. In conclusion, the DDX3 gene-related sex identification of the long-tailed goral appears to be more efficient and precise than the AMEL gene-related approach. This method could be used for the sex identification of the members of the Bovidae family.
본 연구는 한우의 전장유전체 내에 존재하는 단일염기 다형성(SNP)에 대해 DNA chip 분석을 통해 각 개체의 유 전자형을 파악하고, SNP 표지인자간 연관불평형의 크기를 알아보기 위해 실시하였다. 강원도에서 사육되고 있던 한 우 거세우 139두의 조직을 채취하여 DNA를 추출하였으 며, Bovine SNP50K BeadChip 분석을 이용하여 분석에 사 용 가능한 최종 35,769개의 SNP 표지인자를 얻어 연관성 분석을 실시하였다. 분석에 이용된 SNP 표지인자의 총 길 이는 2499.26Mb이었고, 전장 유전체에서 평균 대립유전자 빈도(MAF)는 0.273이었으며, SNP 표지인자간의 평균 거 리는 0.060과 0.082 사이에 분포하였다. 강원지역 내 한우 거세우를 이용하여 확인한 본 실험 결과는 기존의 전국의 한우를 대상으로 한 연구결과와 유사한 패턴을 보였다. 50Kb 이하로 인접한 거리를 갖는 SNP 표지인자들의 연관 불평형 값(r2)이 0.2 초과인 인자가 34%로 나타났다. 또한 SNP 표지인자간 거리가 멀수록 측정값이 떨어지는 지수형 식의 그래프를 보였다. 염색체별로 구분한 40kb 이하로 인 접한 SNP표지인자의 연관불평형 값(r2)은 0.2 이하로 나타 난 것이 총 7개(13, 15, 19, 23, 25, 28, 29번 염색체)였다. 종 합하여 볼 때, 본 연구 결과는 강원한우의 유전적 개량을 이 용한 기초자료로서 활용될 수 있을 것이며, 유전적 개량을 이용한 육종시스템에 도움을 줄 수 있을 것으로 기대된다.
The HNF4α (hepatocyte nuclear factor 4, alpha) is a hepatic transcription factor related to the lipid metabolism and regulation of insulin secretion in humans. The current study about commercial broiler reported that the A543G single nucleotide polymorphism (SNP) within HNF4α gene has an effect on fat deposition and wing yield in the chicken. This study was performed to investigate the association between the SNP within HNF4α gene and growth trait and to verify the applicability as a molecular marker for the improving the performance in Korean native chickens (KNCs). A total of 764 KNCs was collected from the livestock farm of Gyeongnam National University of Science and Technology in Korea and genotyped by PCR-RFLP. The body weight measured at birth, 2, 4, 6, 8, 10, 12, 14, 15, 16, 18, 20, 24, 28, 32, 36, and 40 weeks of age for the association analysis between chicken growth and the SNP genotype. Statistical analysis of the SNP with phenotypic traits was performed using SAS program. The KNC strain was classified by the genotypes of the A543G SNP. The frequencies of three genotypes were 0.47 (AA), 0.43 (AG) and 0.10 (GG), respectively. The SNP of HNF4α has highly significant association (p<0.001) with all-round growth in KNCs. These results suggest that the A543G SNP within HNF4α gene could be a genetic marker for the breeding in Korean native chickens.
The aim of this study is to investigate the genetic interrelationships between economic or meat quality traits (birth weight, weaning weight, average daily gain, market weight, carcass weight, backfat thickness, water, ash, fat, protein, water holding capacity and pH) and 6 SNPs located on six selected candidate genes (MC4R, PGK2, TNNI1, TNNI2, PIK3C3, and CTSK) in Korean native pigs. The genotypes were identified in the 6 SNPs by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) procedure, and association of the genotype on economically important traits was analyzed by general liner model. According to the analyzed results, the MC4R c.1426A>G was correlated with birth weight (p=0.032) and the PGK2 g.122T>G was associated with pH (p=0.026). These findings obtained in the present revealed that the each SNPs of MC4R and PGK2 could be useful as potential genetic markers for birth weight and pH in Korean native pigs, respectively.
Stearoyl-CoA desaturase (SCD1) gene plays important role in fatty acid composition. In order to find marker-assisted selection (MAS) for improving the economic trait, this study was performed to identify the 878T>C single nucleotide polymorphism (SNP) on SCD1 gene. Three genotypes (CC, CT, and TT) were detected in 878T>C SNP of SCD1 gene from 103 Hanwoo population by polymorphism chain reaction-restriction fragment length polymorphism (PCR-RFLP) and economic traits were analyzed by general linear model. The frequency of allele C and T was 0.534 and 0.466, also the genotype frequency of CC, CT and TT was 0.252, 0.563, and 0.184, respectively in the Hanwoo population. The TT genotype of SCD1 gene showed a significantly higher measures (p<0.05) of carcass weight (CW) than CT, CC genotype. No significant association was detected between genotype and other economic trait (marbling, backfat thickness, and longissimus muscle area) in this study. The results revealed that SCD1 gene 878T>C SNP could be useful for effective MAS to increase the economic quality in Hanwoo population.
This study was carried out to investigate artificial insemination (AI) failure status and frozen semen characteristics in Korean proven bulls‘ number (KPN) semen used for AI of Hanwoo cows in Gangwon East region (Gangneung, Donghae, Taebaek, Samcheok, Sokcho, Yangyang, Goseong). Among semen used for AI, AI failure rate showed lowest at KPN506 (27.6%), whereas highest at KPN593 (77.2%). Correlations of AI failure in between Korean proven bulls semen and cows was 0.2941, which means that AI failure rate of Korean proven bulls semen may have respectable effect on reproduction of Hanwoo cow. In addition, present study was conducted to investigate spermatozoal viability rate, ruptured acrosome rate and active mitochondria in frozen Korean proven bulls semen with flow cytometry. The semen of KPN593 showed significantly (p<0.05) higher viability rate in KPN593 (30.49%) than that in KPN637 (37.34%). Furthermore, percentage of ruptured acrosome was lower in KPN637 as 21.37% than in KPN637 (21.37%), but it was not statistically significant. In conclusion, these results indicate that choice of Korean proven bulls semen may correlate positively with conception rate in Hanwoo cow. Therefore, KPN with high AI failure rate might be avoid to increase conception rate and characteristics of frozen semen might be evaluated before its use for AI.
Myopalladin (MYPN) is an important expression gene associated with regulation of Z-line structure in muscle and maintains sarcomeric integrity. In this study, we investigated the association between MYPN A1795G SNP (single nucleotide polymorphism) and carcass traits (LMA, longissimus muscle area; CW, carcass weight; BF, backfat thickness; MS, marbling score) in Korean cattle. The MYPN A1795G SNP was genotyped in 212 steers and analyzed the associations with carcass traits by PCR-RFLP (Restriction fragment length polymorphism) method. The allele frequencies were 0.566 for G allele and 0.434 for A allele. And the genotype frequencies of GG, GA, and AA genotype were 32.1%, 49%, and 18.9%, respectively. Association analysis indicated that the A1795G SNP of MYPN gene showed a significant association with LMA (p<0.05). The steers with GG genotype had higher LMA than those with the genotypes AA. But no significant associations were observed in other carcass traits (CW, BF, MS). The steers with the GG genotype showed higher CW and BF than those with the genotypes AA and GA. These results suggest that the A1795G SNP of the MYPN gene is associated with LMA and may be useful for candidate marker-assisted selection to increase the levels of LMA in Korean cattle.
Enforcement of the beef traceability system can block the sale of imported beef disguised Hanwoo and help customers to know detailed information of the Hanwoo beef while purchase. This study was carried out to investigate the possibilities of identifying DNA upon various ripening and roasting methods (soy sauce, gochujang sauce, 10% vinegar, 10% coke, 10% Cheongha, 10% Soju, completely burned out, raw, normal roasted, dried, and boiled) employed for beef traceability system. Completely burned out meat could not yied useful DNA in the sample, hence wa not analysed. The amplified peaks in most of the samples appeared equally barring completely burned out meat. However, roasted beef after ripened in soy sauce showed new peaks or deletion of a peaks with two markers. The DNA allele height and the amount of extracted DNA in roasted beef after ripening in 10% Cheongha was found to be higher than those in the raw beef. No significant correlations between DNA allele height and the amount of extracted DNA within the samples was noticed. Also, these results suggest that beef traceability using microsatellite (MS) markers is possible in various roasting beef because of bovine specific MS markers. Therefore, beef traceability system by DNA identify test would give the confidence in food safety and beef distribution system to customers.
Analysis of meat quality grade based on the DNA identity test samples were done on 320 Hanwoo (Korean cattle) raised in Gyeonggi region of Korea. 62.29% of cows and 91.35% of steers showed high quality which was above 1st beef grade. However, differences were observed between Gyeonggi Hanwoo and total slaughtered Hanwoo in Gyeonggi in the year 2009, though the sampling was entirely based high quality meat. Meat samples of pedigree registered cattle showed high rate of above 1st meat grade compared to advanced and fundamentally registered cattle. Pedigree registry is associated with quality grade of beef like the sex of cattle. Appearance rate of above 1st beef grade in offsprings of the top 9 Korean Proven Bulls produced through mating to Gyeonggi Hanwoo was 43.27% which was found to be higher than the average grade in cows (47.3%) and was 13.08% higher than the average grade in steers (77.49%). Our results showed superior genetic characters of Korean Proven Bulls which favors and influence the meat quality grade of their offsprings. Furthermore, our results claim that sex of Hanwoo irrespective of pedigreed or Korean Proven Bulls affects and/or influence the meat quality. Conclusively, the results could therefore be used as a basic data for the improvement of beef quality. However, paternity test by DNA typing and huge sample needs to be analysed to authenticate the findings.
Beef traceability, a system that provides all the records of beef production, helps customers purchase that they get to know detailed information of the Hanwoo beef. This study was carried out to investigate the DNA identity possibility in various cooking methods to ripen meat (soy sauce, gochujang sauce, 10% vinegar, 10% coke, 10% Cheongha, 10% Soju, raw, dried, and decayed) for beef traceability. The DNA content of decayed beef was higher than those of other cooking methods. It is thought that result of mixed pollutant, it did not affect amplified DNA allele height because of bovine specific microsatellite (MS) markers. The ripened sample in 10% vinegar 3 days was lowest a mount of extracted DNA (156 ng) and amplified DNA allele height (based on the raw samples to 38%) by MS markers compared with the other cooking methods. There are no significant correlation between amplified DNA allele height and the amount of extracted DNA. Therefore, beef DNA identity test in various cooking methods to ripen meat can used by bovine specific MS markers. Beef traceability system by DNA identify test will give more confidence in food safety to customers
In this study, the analysis of meat quality grade was carried out on the DNA identity test samples of 452 Hanwoo (Korean cattle) raised in Gyeongnam region in the year 2009. Appearance rate of above 1st grade from cow (70%) and steer (93.16%) showed high quality grade. On the other hand, there were the difference between this study and slaughtered Hanwoo in Gyeongnam in the year 2009 - appearance rate of above 1st grade from cow (58.77%) and steer (82.99%). It expect considered influence of the samples for beef DNA identity test that collected mainly high quality meat. Quality grade by registry classification about collected samples showed correlation between pedigree classification and quality grade that appearance rate of above 1st grade highly observed pedigree registry more than advanced registry, advanced registry than fundamental registry. Although limited to DNA identity test samples, KPN (Korean Proven Bull's Number) 452 and KPN 407 was preferred for breeding in Gyeongnam. Also average appearance rate of above 1st grade in offsprings of the top 10 Korean Proven Bulls used mating is 95.35%. It means that Korean Proven Bulls influence meat quality of offsprings positively. Thus, this analysis is expected to be a base data for the improvement of meat quality of Hanwoo raised in Gyeongnam region.