동물조직에서 실시간 유전자 추출을 위한 소형기기의 설계 및 제작

이창근(Chang-Geun Lee),임희택(HeeTaek Lim),윤두학(Duhak Yoon),정일정(Il-Cheong Cheong),정효일(Hyo-Il Jung) 대한기계학회 2008 대한기계학회 춘추학술대회 Vol.2008 No.5

Here, we present a new method and equipment for the extraction of DNA from animal tissues. Our equipment has been designed based on a conventional biochemical analysis system and consists of three major parts; sampling, flow-centrifugation and DNA filtration part. The screw of the sampling part can collect a small piece of tissues from a lump of meat. The piece of meat was subjected to a specially designed tube to extract DNA. The lysed cells mixture were then loaded to the flow-centrifuge part. After the DNA filtration, the elution buffer and cylinder pressure could produce the clean DNA. It is expected in near future that our equipment will help identify the Korean cattle as well as genetically modified organisms.

한우 등심조직 내 succinate dehydrogenase 및 triosephosphate isomerase 발현이 근내 지방함량에 미치는 영향에 관한 연구

Nam-Kuk Kim(김남국),Seung-Hwan Lee(이승환),Dajeong Lim(임다정),Duhak Yoon(윤두학),Chang-Soo Lee(이창수),Oun-Hyun Kim(김언현),Hyeong-Cheol Kim(김형철),Sung-Jong Oh(오성종),Seong-Koo Hong(홍성구) 한국생명과학회 2012 생명과학회지 Vol.22 No.1

비육 전(12개월령)ㆍ후(27개월령)기 한우 등심육을 대상으로 단백체 연구를 통하여 succinate dehydrogenase (SDH) 및 triosephosphate isomerase (TPI) 단백질의 발현 차이가 관찰되었다. 따라서 본 연구는 근내지방함량의 차이를 보이는 비육 전ㆍ후기 한우 등심육 내 차등발현을 보이는 SDH 및 TPI 유전자를 대상으로 근내지방함량과의 관련성 규명을 위하여, 비육 전ㆍ후기 및 한우 동기우 집단 50두를 대상으로 유전자 발현분석 및 통계분석을 수행하였다. 비육 전ㆍ후기 시료를 대상으로 유전자 발현분석을 수행한 결과 SDH 유전자는 12개월령에서 27개월령보다 4배 발현이 높은 것으로 확인되었으며, 한우 동기우 집단 50두 등심육을 대상으로 유전자 발현량과 근내지방함량과의 관련성을 분석한 결과에서도 근내지방함량과 고도의 통계적 유의성(p<0.001)이 있음을 확인하였다. 그러나 TPI의 경우 근내지방함량과의 관련성은 확인되지 않았다. 이러한 결과로 볼 때 SDH 유전자는 한우등심육 내에서 근내지방함량과 관련된 유전자로 판단되며, 지속적으로 유전자구조 변이연구 등을 통한 유전자 마커로의 개발이 필요할 것으로 생각된다. In a previous study, succinate dehydrogenase (SDH) and triose phosphate isomerase (TPI) were detected by 2D gel electrophoresis as differentially expressed proteins in the longissimus thoracis muscles of cattle aged between 12 and 27 months old. In the present study, we investigated the association of SDH and TPI gene expression with intramuscular fat content in 50 Hanwoo steers. The SDH gene was expressed at a 4 times higher level in the 12 month old group than in the 27 month old group (p<0.01). A regression analysis between gene expression value and intramuscular fat content showed a negative correlation between expression of the SDH gene and intramuscular fat content (p<0.001). In contrast, the expression of the TPI gene showed no significant association with intramuscular fat content. This result suggests that the SDH gene may be a candidate marker gene for intramuscular fat content in the longissimus thoracis of Korean cattle.

한우 등심부위 근육 내 조지방함량에 따른 inducible cAMP early repressor (ICER) 유전자발현 분석

이승환(Seung Hwan Lee),김남국(Nam Kuk Kim),김성곤(Sung Kon Kim),조용민(Yong Min Cho),윤두학(Duhak Yoon),오성종(Sung Jong Oh),임석기(Seok Ki Im),박응우(Eung Woo Park) 한국생명과학회 2008 생명과학회지 Vol.18 No.8

근내지방 축적이 왕성하게 진행되는 12개월 및 27개월령 사이에 유전자발현이 증가하는 inducible cAMP early repressor(ICER) 유전자의 클로닝 및 유전자발현 특성을 규명하기 위하여 한우조직별, 근육 내 조지방 함량에 따라 마블링 high- 및 low 한우 각 4두와 3두씩을 공시하였다. ddRT-PCR 분석을 통하여 증폭된 300 bp PCR 산물 및 EST 서열을 중심으로 1.5 kb 한우 ICER cDNA를 염기서열 분석하였으며, 조직별 유전자발현분석결과, 식이지방이 흡수되는 소장 조직에서 ICER 유전자의 발현량이 가장 높았다. 아울러, 같은 근육타입인 등심 및 우둔조직 사이에서 ICER 유전자발현은 등심조직에서 2.5배 많이 발현하였다. 한우 마블링 high, low 두 그룹 간 유전자발현 분석 결과, high 그룹에서 ICER 유전자발현이 4배 이상 증가하는 것으로 분석되었다. 따라서, ICER 유전자는 가축의 마블링과 밀접한 관련이 있는 유전자임이 사료된다. Marbling (intramuscular fat) is an important factor in determining meat quality in Korean beef market. A grain based finishing system for improving marbling leads to inefficient meat production due to an excessive fat production. Identification of intramuscular fat-specific gene might be achieved more targeted meat production through alternative genetic improvement program such as marker assisted selection (MAS). We carried out ddRT-PCR in 12 and 27 month old Hanwoo steers and detected 300 bp PCR product of the inducible cAMP early repressor (ICER) gene, showing highly gene expression in 27 months old. A 1.5 kb sequence was re-sequenced using primer designed base on the Hanwoo EST sequence. We then predicted the open reading frame (ORF) of ICER gene in ORF finder web program. Tissue distribution of ICER gene expression was analysed in eight Hanwoo tissue using realtime PCR analysis. The highest ICER gene expression showed in Small intestine followed by Longissimus dorsi. Interestingly, the ICER gene expressed 2.5 time higher in longissimus dorsi than in same muscle type, Rump. For gene expression analysis in high- and low marbled individuals, we selected 4 and 3 animal based on the muscle crude fat contents (high is 17-32%, low is 6-7% of crude fat contents). The ICER gene expression was analysed using ANOVA model. Marbling (muscle crude fat contents) was affected by ICER gene (P=0.012). Particularly, the ICER gene expression was 4 times higher in high group (n=4) than low group (n=3). Therefore, ICER gene might be a functional candidate gene related to marbling in Hanwoo.

국내 양돈장의 사육구간별 주요 소화기질병 원인체 유병율 조사

정윤수 ( Youn-soo Jung ),박유리 ( Yu-ri Park ),강대영 ( Dae-young Kang ),한도현 ( Do-hyun Han ),윤두학 ( Duhak Yoon ),정병열 ( Byeong-yeal Jung ),박최규 ( Choi-kyu Park ) 한국가축위생학회 2016 韓國家畜衛生學會誌 Vol.39 No.4

For determining the prevalence of major enteric pathogens, clinical examination and etiological diagnosis were carried out on 75 Korean pig farms. Enteric disease-suspected signs were observed in 90.7% of the farms and the incidence and severity were higher in younger age groups of the pigs. Five of seven pathogens were detected in 375 fecal samples collected from the 75 farms, and the farm-level prevalence of porcine rotavirus group A (PoRVA), pathogenic Escherichia (E.) coli, Lawsonia (L.) in-tracelluraris, Salmonella spp., and Brachyspira (B.) hyodysenteriae was 54.7%, 54.7%, 16.0%, 10.7% and 2.7%, respectively. PoRVA was extensively infected in suckling and weaning pig groups. The prevalence of pathogenic E. coli was highest in suckling period, and after the period, it exhibited a tendency to decrease. Salmonella spp. and L. intracelluraris were detected in all feeding groups of pigs in a ratio of 1.3∼6.7%. B. hyodysenteriae was detected in 1.3∼2.7% of growing and fattening pig groups but not detected in suckling and weaning pig groups. At least one or more pathogens were detected in 30.1% of 375 fecal samples. Among these, 25.0% or 5.1% of cases were single or mixed infection. Enteric disease signs of the pigs were significantly co-related with the detection of PoRVA, pathogenic E. coli or Salmonella spp. (P<0.01) but not with L. intracelluraris or B. hyodysenteriae (P>0.05). Conclusively, it will be expected that these data obtained in this study are very useful for subsequent studies and prevention strategies for swine enteric disease in Korean pig farms.

전장 유전체 관련성 분석을 통한 한우 도체수율 관련 양적형질좌위 탐색

이승환,임다정,당창권,장선식,김형철,전기준,연성흠,장길원,박응우,오재돈,이학교,이준헌,강희설,윤두학 충남대학교 농업과학연구소 2013 농업과학연구 Vol.40 No.2

Genome-wide association study was performed on data from 266 Hanwoo steers derived from 66 sire using bovine 10K mapping chip in Hanwoo (Korean Cattle). SNPs were excluded from the analysis if they failed in over 5% of the genotypes, had median GC scores below 0.6, had GC scores under 0.6 in less than 90% of the samples, deviated in heterozygosity more than 3 standard deviations from the other SNPs and were out of Hardy-Weinberg equilibrium for a cutoff p-value of 1-15. Unmapped and SNPs on sex chromosomes were also excluded. A total of 4,522 SNPs were included in the analysis. To test an association between SNP and QTL, GWAS for five genetic mode(additive, dominant, overdominant, recessive and codominant) was implemented in this study. Three SNPs (rs29018694, ss46526851 and rs29018222) at a threshold p<1.11×10-5 were detected on BTA12 and BTA21 for dressing percentages in codominant and recessive genetic mode. The G allele for rs29018694 has 4.9% higher dressing percentage than A allele, while the T allele for ss46526851 has 2.57 % higher dressing percentage than C allele. Therefore, rs29018694 SNP showed a bigger effect than the other two SNPs (ss46526851 and rs29018222) in this study. In conclusion, this study identifies three loci with moderate effects and many loci with infinitesimally small effect across genome in Hanwoo.