Earticle

PMO-P4, being cultivated as “Sanghwang”in Korea, was proved to be P. baumii based on ITS (internal transcribed spacer) sequencing and RFLP (Restriction Fragment Length Polymorphism) patterns along with some Phellinus species including P. linteus. The similaraty of ITS sequencing between PMO-P4 and other Phellinus species was given the range of 48.6%~72.2%, showing the highest homology from P. linteus and the lowest from P. gilvus

The fatty acid binding protein 4 (FABP4) gene plays an important role in lipid metabolism and homeostasis in adipocytes. The objective of this study was to analyze the association between a single nucleotide polymorphism (SNP), g.7516G>C, in the FABP4 gene and economic traits of Korean native cattle, Hanwoo. Primers were designed to target a region of the FABP4 gene between nucleotides 7417 and 7868 (AAFC01136716). The SNP, which was detected by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method using restriction enzyme MspA1I, was genotyped in 319 animals of Hanwoo steer population. Statistical analysis showed that the SNP genotype of the FABP4 gene significantly affected carcass weight (CW, p<0.01), longissimus muscle area (LMA, p<0.001), and marbling score (MS, p<0.001). GG allele of the SNP on 246 animals in a Wagyu × Limousin F2 reference population showed a higher MS (p<0.05) and subcutaneous fat depth (p<0.05) in previous report. But CC allele of the SNP showed greater values for MS, LMA, and CW in Hanwoo steers. These results suggest that the g.7516G>C SNP located in the FABP4 gene may affected differently depending on the cattle breed and can be used as a genetic selection marker in Korean native cattle.

The role of Fatty acid binding protein 4 (FABP4) gene is critical for lipid metabolism and for maintaining homeostasis in adipocytes. Association between Hanwoo carcass traits and FABP4 gene g.7516G>C SNP has been reported previously, however, its association and how does it influence Korean brindle and black cattle has not been demonstrated and established till date. For this purpose, the study was planned to analyze the SNP association (Single Nucleotide Polymorphism) (g.7516G>C) in FABP4 gene and gather genetic information on economic traits of Korean brindle cattle and black cattle. As per sequence of bovine FABP4 gene available (Genbank accession No. NC_007312.4), one pair of primers (5’-ATA TAG TCC ATA GGG TGG CAA AGA-3’ and 5’-AAC CTC TCT TTG AAT TCT CCA TTC T-3’) was designed to amplify a 452bp product of the FABP4 gene including the region of 7417–7868. The SNP, detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) methods using restriction enzyme MspA1I, was genotyped in 117 animals of brindle cattle and 24 animals of black cattle population. Statistical analysis revealed that the FABP4 genotype significantly (p<0.05) affect with carcass weight, but there was no significant association with any other economic traits was observed in brindle as well as in black cattle. In conclusion, these results suggested that SNP (g.7516G>C), located in FABP4 gene, could be used as important DNA marker of economic traits in Korean brindle cattle. Furthermore, we suggest that additional samples needs to further analysed to make related data exclusively authentic.

This study was conducted to detect the specific fragment genes by using RAPD-PCR and RFLP method in the Korean Tiger cattle and Korean Native cow. And then, the specific fragment gene was investigated by the analysis of the genes for detection significance according to the expressing pattern. We found the specific expression gene by the RAPD-PCR analysis in Korean Tiger cattle. It were a detected the differences of the species in the colour and external section. The Korean Tiger cattle were vary low compare to the Korean Native cattle by analysis result of polymorphism and distribution. And the polymorphism over 500bp into the size classification detected was highly pattern and it could be utilize by the resource of the specific gene. There was a found the specific gene by sequencing in the 1855bp gene fragment of Korean Tiger cattle. And the sequencing result of the R9B was different between Korean Native cow and Holstein cattle. Thus, this gene can be apply as the specific gene in the Korean Tiger cattle.

The objective of this study was to find out candidate genes associated with litter size trait in pigs of inbred Large Yorkshire and Landrace populations. 86 sows were screened for candidate genotypes along with litter size data recordings. Association of litter size with genotypes of candidate genes were investigated to verify the usefulness of each gene's genotypes as markers for the trait. For the lines of Large Yorkshire, PRLR3 and RBP4 genes were genotyped. Frequency distribution of PRLR3 with genotypes AA, AB and BB were each 0.14, 0.44 and 0.42. And the average litter size by PRLR3 genotypes were 8.83, 10.81 and 10.70 piglets per litter, the average estimated breeding values of which were 0.243, 0.332, 0.365, respectively for AA, AB and BB genotypes. Genotypic frequencies of RBP4 by AA, AB and BB genotypes were 0.10, 0.44 and 0.46. The average litter size by genotypes of RBP4 were 10.40, 10.57 and 10.35 piglets per litter and their corresponding average estimated breeding values were 0.451, 0.353 and 0.261, respectively for genotypes AA, AB and BB. Significance in differences among genotypes were not observed, but B allele of RBP4 seems to be associated with litter size. In Landrace lines, frequencies of RBP4 genotypes, AA, AB and BB were 0.29, 0.55 and 0.16. And the average litter size of these genotypes were 10.50, 11.08 and 11.00 piglets per litter. The corresponding averages of estimated breeding values of each genotypes were 0.172, 0.135 and 0.104. In Landrace lines, allele A was more likely to be associated with litter size, even if differences among average litter size were not significant. We conclude that genotyping of two candidate genes is a helpful tool to identify genetic potentials of litter size in pigs.

The relationship of several candidate genes polymorphisms with breeding values of economic traits were investigated in Korean Native Pigs. Record (2001-2006) of 546 Korean native pigs were obtained from National Institute of Animal Science, Korea having data on average daily gain (ADG), age at 70 kg (D70 kg) and backfat thickness (BF). The data's obtained were analyzed by the DF-REML (Derivative-Free Restricted Maximum Likelihood) program of Boldman using a single-trait animal model to analyse the genetic parameters. The analysis of restriction fragment length polymorphism (RFLP) was conducted on 68 Korean native pigs (KNP) using single nucleotide polymorphism (SNP). Different genotype frequencies of 5 candidate genes such as MC4R, PRKAG3, FABP3, ESR and PRLR3 were observed in KNP. Significant relationship of AA and AB genotype between MC4R polymorphic site and breeding value for average daily gain (ADG, p<0.05) was observed. PRKAG3 polymorphic sites were also found to be significantly related to breeding values for ADG, AA, AB genotype (p<0.05) and also, for Backfat thickness (BF), days to 70 kg and BB genotype (p<0.05). In conclusion, selection method would be more effective if it encompasses significant genotype for performance traits and that would further aid in the selection of seed stock in KNP.

본 연구는 도체성적을 보유하고 있는 제 31차, 32차 한우 후보종모우 집단 228두를 선발 하여 DNA를 분리 정제 후 exon 2에 위치한 bovine leptin gene 염기서열 가운데 특정 염기서열을 갖는 2 좌위의 primer를 합성하여 PCR 수행 후 PCR-product를 이용하여 2 종류의 제한효소 Kpn 2 I, Msp I 으로 반응시킨 후 두 가지 형태의 대립 유전자를 검정하여 경제형질과의 관련성을 분석하였다. PCR-RFLP를 통하여 얻어진 leptin gene의 유전자형 빈도는 Kpn2 I의 경우 C 유전자 빈도(0.25)보다 T 유전자 빈도(0.75)가 높게 나타났으며 Msp I으로 처리한 경우 M 유전자 빈도(0.35)보다 m 유전자 빈도(0.65)가 높게 나타났다. 통계적 분석을 통하여 각 유전자형에 대한 경제형질과의 관련성을 분석한 결과 제한효소 Kpn2 I으로 처리한 경우 도체율에서 CT 유전자형과 CC 유전자형 사이에 유의적 차이가 나타났으며(p<0.05), Msp I의 경우 도체중 Mm 유전자형과 mm 유전자형 사이에 통계적 유의성이 나타났다(p<0.05).

The identification of the leptin gene in 1994 and it's adipocytes specific protein leptin has provided the first physiological links to the regulatory system controlling body weight and fat deposits. The meat tastes is mainly determined by quantify and quality of triglyceride stored in adipose tissue. This study was conducted to analyze genetic characteristics of Hanwoo leptin gene and also to investigate the association of DNA marker with some economic meat traits for Hanwoo.The leptin hormone gene polymorphisms were identified by digestion with Kpn2Ⅰ and MspⅠ. Slaughter weight(SWT), slaughter percentage(SP), longissimus muscle area(LMA), beef marbling score(MS) and back fat thickness(BF) were compared among three genotypes by PCR-RFLP and showed significant differences among genotypes. PCR-RFLP(Kpn2Ⅰ) were detected significant for SP, MS and BF. The allele was associated with fatter carcasses and C allele with leaner carcasses.

본 연구는 Pvu II PCR-RFLP를 이용하여 Yorkshire종 돼지에서 Estrogen Receptor의 유전적 다형과 산자수간의 관련성을 분석하기 위하여 수행되었다. 무창 돈사에서 사육중인 242두의 종빈돈으로부터 혈액을 채취하여 Pvu II PCR- RFLP로 ER 유전자형을 결정하였다. ER 유전자 좌위에서 유전자 빈도는 각각 0.39(A)와 0.61(B)였다. ER 유전자형별 산자수에 대한 효과는 최소제곱평균을 설명하는 고정모형을 설정하여 추정하였다. 분석결과 복당 총산자수와 실산자수에서 특정 ER 대립유전자(B)에서 산자수 증진효과가 관찰되었다. 따라서 돼지 ER 좌위의 유전적 변이는 번식돈의 산자수 증대와 관련된 marker-assisted selection(MAS)에 응용될 수 있을 것으로 사료된다.

This study was performed to investigate a possible association of the porcine estrogen receptor(ER) locus with the total number of born(TNB) and number of born alive(NBA) in Yorkshire pigs. Using DNAs extracted from 242 Yorkshire pigs, the ER genotype was determined by Pvu II PCR-RFLP. The ER allele frequencies of two types of A and B were 0.39 and 0.61, respectively. The least squares means of the litter size by ER genotype was evaluated. The TNB and NBA were found to be associated with an specific ER allele. The genotype at the porcine ER locus has an application potential for marker-assisted selection for litter size in pigs.

This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.

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.

Because the import of Inonotus obliquus have been rapidly increased in Korea, we developed the Inonotus species-specific marker by using various sequences including ITS sequences, PCR-RFLP and STS primers and used this marker to determine both genetic relatedness and strains discrimination of Inonotus spp. Total 17 different Inonotus spp. were examined by using ITS sequences and classified into 2 different groups. One strain, ASI74008 isolated from Kamchaka island of Siberia, showed the high sequence identity (98%) at the nucleotide level to the other I. obliquus DSM strain, indicating the ASI74008 belong to I . obliquus species. Comparison of banding patterns after restriction enzyme digestions with PCR amplicons of ITS region revealed some variations depending on the species and strains. However, PCR products amplified with STS primer showed species specific patterns.Therefore, use of both STS primers and PCR-RFLP could help for better strain identification.

최근 재래가축 유전자원의 중요성이 부각되면서 그 중의 칡소의 유전형 확립이 대두 되고 있다. 칡소는 호반모를 가진 종모우의 정액을 이용하여 수정하여도 호반모가 아닌 일반 한우의 모색과 비슷한 양상으로 출현하는 개체들이 있다. 이는 칡소 모색 관련 유 전자들의 발현이 고정되지 않았다는 가능성을 제시한다. 소의 모색관련 유전자 중에서 melanocortin 1 receptor(MC1R) 유전자의 결실은 한우의 모색 및 한우고기의 판별에 이 용되어 왔다. 본 연구는 PCR-RFLP 기법으로 melanocortin 1 receptor(MC1R) 좌위의 유 전자형 검출을 통해 칡소와 한우의 유전자형 빈도와 염기 서열상의 차이를 비교 분석함 으로써, 칡소의 모색 발현에 관련된 MC1R 유전자를 이용하여 호반모 발현 비율과 유전 양상을 밝히기 위하여 수행되었다. 본 연구는 전라북도 축산위생연구소 축산시험장에서 사육 중인 칡소로 부계나 모계를 알고 있거나 가계가 형성된 칡소로 부터 혈액을 채취하여, Genomic DNA를 추출하였다. MC1R 특이 유전자 마커를 이용하기 위해 GenBank(S71017)에 등록된 염기 서열을 참고 하여 Primer를 설계하였다. MC1R 유전자의 증폭을 위한 PCR 반응 조건은 95℃에서 10 분간 정치 후 95℃ 30초, 60℃ 30초, 72℃ 1분 총 35 cycle을 수행한 뒤 72℃에서 5분 간 반응시켜 PCR을 완료하였다. 증폭된 MC1R 유전자 739 bp산물을 MspI으로 3시간 처 리 후 전기 영동하여 DNA 절편을 확인하였다. PCR 증폭 산물을 MspI 처리한 결과, 한우의 절편 양상은 2개의 절편(534 bp, 207 bp) 으로 대부분 나타났으며, 칡소는 4개의 절편(535 bp, 328 bp, 207 bp, 174 bp) 양상이 3 개의 절편(328 bp, 207 bp, 174 bp) 양상보다 다소 많은 경향을 보였다. 이 연구에서 얻 어진 한우와 칡소의 유전자 절편 양상을 비교 분석하여 호반모 발현비율을 증가시키는 교배체계의 확립에 이용될 수 있을 것으로 기대된다.

Pleurotus has increased rapidly production and consumption because of highly nutritional value, natural healthy food and so on. The basic studies for Pleurotus need for development of mushroom industry. This study was to investigate the cultural characteristics among 15 strains of 6 species and to analyze their phylogenetic relationships. The cultural characteristics were investigated by mycelial growth activity at different media, temperature and pH. The optimum media for mycelial growth were YM and MCM in most species. The optimum temperature for mycelial growth were 25oC and 30oC. The optimum pH for mycelial growth were widly range from pH 5.1 to 7.4. Through the RFLP (restriction fragment length polymorphism) of IGS (intergenic spacer) I region in ribosomal DNA, it was analyzed phylogeny of interspecies and intraspecies. Each species was discriminated well as isolates within each species formed clade to be distinguished other species. P. florida was highly similar to P. floridanus, and P. flabellatus was P. cornucopiae. P. fuscus var. ferulae was highly similar to P. eryngii but discriminated different species in analysis of RFLP of IGS I region and showed different characteristics in mycelial culture. RFLP of IGS I region was useful of studying phylogenetic relationships of species and population.

Genomic DNA prepared from fruitbdies of 25 Grifola frondosa strains were amplified with the ITS primers and the PCR reaction products were enzymed. The ITS bands have same electrophoresis patterns but part of them have different restriction enzyme cutting site. These strains were divided into three broad categories. SRAP amplification employing 47 SRAP primer pairs were carried on and 138 polymorphic bands were detected. The phylogram tree showed that: ten strains were differentiated from the others effectively and fifteen strains have no obvious differences between each other. The results implied that the ITS-RFLP and SRAP markers were effective methods for strains identifica tion and germplasm evaluation but other markers were also needed to be used in conjunction.

본 연구는 319두의 서로 다른 품종에서 PSE육을 생산하는 PSS 돼지 출현빈도를 조사하였다(Yorkshire 150; Landrace 89 and Duroc 80). PCR-RFLP법을 이용하여 돼지의 모근을 DNA sample로 사용하여, PCR로 증폭된 유전자는 Cfo I 제한 효소로 절단하여 종돈에 존재하는 ryanodine receptor (RYR 1) 돌연변이 유전자의 출현빈도를 조사한 결과를 요약하면 다음과 같다. 모근에서 추출한 DNA를 주형으로 한 Primary PCR을 수행한 결과 ryanodine receptor 유전자 중 659bp의 증폭산물을 얻었으며, second PCR을 수행한 결과에서는 522 bp의 증폭산물을 얻었다. 이 증폭산물은 porcine ryanodine receptor 유전자의 exon 영역 중 PSS를 유발하는 point mutation(C→T; Arg→Cys) 부분을 포함하고 있으므로 Cfo I 제한효소에 의해 분석될 수 있으며, agarose gel 전기영동에 의하여 세 가지의 유전자형으로 분류할 수 있다. 정상 homotype(NN)은 두 개의 DNA band(439, 83bp)로 나타나며, 열성 homotype(nn)은 552 bp의 단일 밴드로 출현한다. 그리고 세 개의 밴드(522, 439 그리고, 83 bp)는 heterotype(Nn)의 잠재성 돼지로 표현된다. Yorkshire종에서는 정상돼지가 98.00%로 나타났으며, hetero 돼지는 2.00% 그리고, PSS돼지는 출현하지 않았다. Landrace 돼지에서는 정상돼지가 87.64%로 나타났으며, hetero 돼지와 PSS패지가 각각 11.24와 1.12%로 나타났으나, Duroc종에서는 정장돼지(NN)만이 출현하였다. 대립 유전자 빈도는 Yorkshire종은 정상 N유전자가 0.990의 비율로 나타났으며, 열성 n 유전자는 0.010의 비율로 출현하였으며, Landrace종에서는 N유전자와 n유전자가 각각 0.933과 0.067의 빈도로 출현하였으며, Duroc종에서는 N 유전자의 빈도가 1.000의 빈도로 나타났으나, n유전자의 빈도는 0.000의 빈도로 나타났다. 3품종 집단 모두에서 Hardy-Weinberg 법칙과 일치하여 유전적 평형을 이루고 있었다.

This study was carried out to investigate PSS (Porcine Stress Syndrome) with the PSE (Pale, Soft, Exudative) in 319 different pigs(Yorkshire 150; Landrace 89 and Duroc 80). The PCR-RFLP method was adapted to detect the ryanodine receptor (RYR 1) gene mutation and to estimate the genotype frequency of the RYR1 gene in breeding pig population. The DNA samples were collected from hair follicles of pigs of Yorkshire, Landrace and Duroc. After DNA amplification by PCR, the PCR products were digested by restriction enzyme, Cfo I. Primary PCR products of ryanodine receptor gene were length of 659 bp in hair follicle and their second PCR products were length of 522 bp in hair follicle. The exon region (522 bp) including point mutation (C→T; Arg→Cys) in the porcine ryanodine receptor gene, which is a causal mutation for PSS, was digested with Cfo I restriction enzyme. The RYR1 gene was classifed into three genotypes by agarose gel electrophoresis. The normal homozygous (NN) individuals showed two DNA fragments consisted of 439 and 83 bp. The mutant homozygous (nn) individuals showed only one DNA fragment 522 bp. In addition, all three fragments (522, 439 and 83 bp) were showed in heterozygous (Nn) carrier animals. The normal homozygous (NN), heterozygous (Nn) and mutant homozygous (nn) were 98.00, 2.00 and 0.00% in Yorkshire pigs, 87.64, 11.24 and 1.12% in Landrace, 100.00, 0.00 and 0.00% in Duroc, respectively. The gene frequencies of N and n were 0.990 and 0.010 in Yorkshire pigs, 0.933 and 0.067 in Landrace, 1.000 and 0.000 in Duroc, respectively.