• Asian-Australasian Journal of Animal Sciences
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• 제27권9호
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• pp.1254-1262
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• 2014

Lymphoid enhancer binding factor 1 (LEF-1) is a member of the T-cell specific factor (TCF) family, which plays a key role in the development of breast endothelial cells. Moreover, LEF-1 gene has been identified as a candidate gene for teat number trait. In the present study, we detected two novel mutations (NC_010450.3:g. 99514A>G, 119846C>T) by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism in exon 4 and intron 9 of LEF-1 in Guanzhong Black, Hanjiang Black, Bamei and Large White pigs. Furthermore, we analyzed the association between the genetic variations with teat number trait in these breeds. The 99514A>G mutation showed an extremely significant statistical relevance between different genotypes and teat number trait in Guanzhong (p<0.001) and Large White (p = 0.002), and significant relevance in Hanjiang (p = 0.017); the 119846C>T mutation suggested significant association in Guanzhong Black pigs (p = 0.042) and Large White pigs (p = 0.003). The individuals with "AG" or "GG" genotype displayed more teat numbers than those with "AA"; the individuals with "TC" or "CC" genotype showed more teat numbers than those with "TT". Our findings suggested that the 99514A>G and 119846C>T mutations of LEF-1 affected porcine teat number trait and could be used in breeding strategies to accelerate porcine teat number trait improvement of indigenous pigs breeds through molecular marker assisted selection.

• 한국육종학회지
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• 제42권5호
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• pp.515-524
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• 2010

본 연구에서는 RNAi 기작을 이용하여 식미에 중요한 영향을 미치는 아밀로스 함량을 다양화하기 위해 GBSSI 유전자의 3'-UTR 부위를 targeting하여 dsRNA를 생성시킬 수 있는 운반체를 제작하고, 벼에 형질전환 하였다. 작성된 형질전환체들을 대상으로 $I_2$-KI 용액 반응과 아밀로스 함량을 분석한 결과, $I_2$-KI 용액에 대한 반응은 waxy 타입으로 나타났으나 아밀로스 함량은 찰벼와 저아밀로스 벼 사이에 해당되는 범위를 보였다. 원품종과 형질전환체 간의 아밀로펙틴 사슬 분포의 차이를 비교한 결과, 단쇄에 해당되는 A1과 B1 사슬의 분포는 감소한 반면, 중쇄에 해당되는 B2와 장쇄인 B3 사슬의 분포는 다소 증가하였으며, B3 사슬의 분포비율은 사용된 품종에 따라 약간의 차이를 보였다. 배유 단면의 전자현미경적 구조를 비교한 결과, 원품종에 비해 형질전환체의 전분립 크기가 작아지고 쪼개짐의 형태가 완만한 굴곡을 보였다. 이러한 결과를 바탕으로, RNAi 기술을 이용하여 다양한 아밀로스 함량이 조절된 형질전환 벼를 개발하기 위해서는 targeting 부위를 결정하는 것이 하나의 중요한 전략이 될 수 있음을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제28권1호
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• pp.20-24
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• 2015

Insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-1 (IGFBP-1) play a pivotal role in regulating cellular hypoxic response. In this study, we cloned and characterized the genes encoding IGF-1 and IGFBP-1 to improve the current knowledge on their roles in highland Bos grunniens (Yak). We also compared their expression levels in the liver and kidney tissues between yaks and lowland cattle. We obtained full-length 465 bp IGF-1 and 792 bp IGFBP-1, encoding 154 amino acids (AA) IGF-1, and 263 AA IGFBP-1 protein, respectively using reverse transcriptase-polyerase chain reaction (RT-PCR) technology. Analysis of their corresponding amino acid sequences showed a high identity between B. grunniens and lowland mammals. Moreover, the two genes were proved to be widely distributed in the examined tissues through expression pattern analysis. Real-time PCR results revealed that IGF-1 expression was higher in the liver and kidney tissues in B. grunniens than in Bos taurus (p<0.05). The IGFBP-1 gene was expressed at a higher level in the liver (p<0.05) of B. taurus than B. grunniens, but it has a similar expression level in the kidneys of the two species. These results indicated that upregulated IGF-1 and downregulated IGFBP-1 are associated with hypoxia adaptive response in B. grunniens.

• 대한임상검사과학회지
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• 제55권3호
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• pp.184-194
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• 2023

골격근은 대사, 열기반 온도 조절, 그리고 전반적인 체내 균형을 위해 필수적인 조직이고 근발생(myogenesis)이라는 다단계 과정을 거쳐서 근관세포를 형성한다. p-아니스알데하이드(p-anisaldehyde, PAA) (4-메톡시벤잘데하이드)는 아니스 씨에서 추출된 에센셜 오일의 주성분이지만, 골격근에서의 기능은 아직까지 알려져 있지 않다. 따라서, 우리는 마우스 C2C12 근육모세포를 이용하여 근육분화가 PAA에 의해 영향을 받는지를 연구하였다. C2C12 근육모세포의 분화를 유도하기 위해 이 세포를 분화배지에서 5일동안 배양하였고, 매일 PAA (50 또는 200 ㎍/mL)를 포함하는 새로운 배지로 교체하였다. 대조군으로서 PAA가 포함되지 않은 배지를 사용하였다. 우리는 분화시작 후 1, 3, 5일째에 근관세포의 길이와 지름을 측정함으로써 PAA가 근관 형성에 미치는 영향을 평가하였고, quantitative real-time polymerase chain reaction 분석을 통해 PAA가 근육 표지인자(myoblast determination protein 1, myogenin, myocyte enhancer factor 2C, muscle creatine kinase, 및 myosin heavy chain)와 근육위축 관련 유전자(atrogin-1과 muscle ring finger-1 [MuRF-1])의 발현에 미치는 영향을 분석하였다. 또한, 주요 근육형성 키나아제인 protein kinase B (Akt)의 인산화를 웨스턴 블롯을 이용해 관찰하였다. 그 결과 PAA가 더 작고 얇은 근관 형성을 유의하게 유발하며 근육 표지인자의 발현을 감소시킨다는 것을 확인하였다. 또한, atrogin-1과 MuRF-1의 발현이 PAA에 의해서 감소하였는데, 이는 Akt 인산화의 감소와 일치하는 결과이다. 결론적으로, 본 연구결과는 PAA가 Akt 인산화와 활성화를 감소시킴으로써 C2C12 세포에서의 근육 분화를 억제하는 역할을 한다는 것을 증명한다.

• Toxicological Research
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• 제19권2호
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• pp.115-121
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• 2003

In view of the importance of muscle-specific creatine kinase (CKMM) gene as a genetic factor for athletic performance, we investigate the relationship between elite athletic performance and two restriction fragment length polymorphisms (Ncol and Taql RFLPs) in the CKMM gene. Genomic DNA was extracted from white blood cells of 98 unrelated male Korean elite athletes and 04 sedentary controls, respectively. Two genetic polymorphisms in the CKMM gene were detected by the polymerase chain reaction and the digestion with restriction endonucleases, Ncol and Taql, respectively. There were no significant associations between two genetic polymorphisms in the CKMM gene and elite athletic performance or clinical parameters in our subjects. Therefore, these findings suggest that two genetic polymorphisms in the CKMM gene may not be useful as genetic markers to predict the athletic performance in male Koreans.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.178-186
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• 1995

Streptomyces albus wild type ATCC 21838 produced salinomycin, polyether antibiotic. To clone genes related salinomycin production, a genomic library was screened using actI as a DNA hybridization probe. pWHM 210 was isolated, which contained an approximately 24 kb of insert DNA. A 3.8 kb region in the 24 kb insert DNA was hybridized to actI and the nucleotide sequence of this region was determinied. Two open reading frames found in the same direction were homologous to genes for $\beta$-keto acyl synthase/acyl transferase and chain length determining factor in type II PKS (polyketide synthase). The genes were components of minimal type II PKS genes, highly conserved and showed the strong simiarity to other type II PKS genes known today.

• 대한의생명과학회지
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• 제29권3호
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• pp.190-200
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• 2023

Skeletal muscle, essential for metabolism, thermoregulation, and immunity, undergoes myogenic differentiation that results in myotube formation. Trans-anethole (TA), the major constituent in essential oil produced by anise, star anise, and fennel, whose function in skeletal muscle has not yet been elucidated. Therefore, we investigated whether TA influenced muscle differentiation in mouse C2C12 myoblasts. Cells were induced to differentiate using a differentiation medium with or without TA (50 or 200 mg/mL) daily for 5 days. We measured myotube length and diameter after differentiation days 1, 3, and 5 and analyzed the expression of myogenic markers (myoblast determination protein 1, myogenin, myocyte enhancer factor 2, muscle creatine kinase, and myosin heavy chain) and atrophy-related genes (atrogin-1 and muscle ring finger-1 [MuRF-1]) using quantitative real-time PCR. Additionally, we observed the expression of total protein kinase B (Akt) and phosphorylated Akt (p-Akt) using western blotting. Our data showed that TA significantly induced the formation of smaller and thinner myotubes and reduced the myogenic factor expression. Furthermore, the atrogin-1 and MuRF-1 expression markedly increased by TA. Consistent with these findings, TA significantly decreased the expression of total Akt and p-Akt. Taken together, these results indicate that TA inhibits myogenic differentiation of C2C12 cells via reduction of both total Akt and p-Akt. Our findings may provide valuable insights into the impact of PAA on individuals at risk of muscle atrophy.

• 한국해양바이오학회지
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• 제1권2호
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• pp.119-125
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• 2006

어류의 insulin-like growth factor-I (IGF-I)의 생화학적 작용기작을 연구하기 위하여 한국산 조피볼락의 IGF-I cDNA 유전자 cloning을 행하였다. 완전한 cDNA 유전자 염기서열은 PCR과 RACE 방법을 통하여 얻어진 DNA로부터 결과를 얻을수 있었다. 결정된 IGF-I의 염기서열은 flounder, chinook salmon, human IGF-I의 염기서열과 비교한 결과 각각 93.6%, 90.7%, 85.4%의 높은 상동성을 보였다. 생화학적으로 활성이 있는 재조합 IGF-I을 얻기 위하여 IGF-I의 B-C-A-D domain 부분을 PCR로 얻은 뒤 E. coli BL21(DE3)에 넣어 overexpression 시켰다. Ni-NTA colummn을 사용하여 순수한 재조합 단백질을 정제할수 있었다. 정제된 단백질은 SDS-PAGE 상에서 7 kDa의 단일 band를 보여 주었으며 [$^3H$]-thymidine 결합정도를 측정하는 방법으로 활성을 가지고 있음을 확인할수 있었다.

• 미생물학회지
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• 제33권4호
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• pp.237-241
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• 1997

Streptomyces coelicolor A3(2) 배양액의 lipase(EC 3.1.1.3)는 ${\alpha}$-naphthyl-butyrate에 대하여 활성을 나타내어 ${\alpha}$-naphthyl-acetate에 활성을 나타내는 esterase와 구분할 수 있었다. Streptomyces coelicolor A3(2) 세포외 lipase를 Sephadex G-100, DEAE-Cellulose 그리고 Phenyl-Sepharose CL4B 크로마토그래피의 과정을 통해 16% 수율로 15배 분리정제 하였다. SDS-폴리아크릴아마이드 겔 전기영동에서는 34.7 kDa정도의 분자량을 갖는 것으로 나타났다. Tributyrin를 기질로 사용하였을 때 이 lipase의 최적활성조건은 pH 8에서 9 그리고 $37^{\circ}C$였다. 기질로서 triacylglycerol의 지방산 길이가 증가할수록 활성은 감소하였다. A-factor에 의해 lipase활성이 억제되므로 배양초기의 낮은 lipase활성과 관련될 것으로 보인다.

• Animal Bioscience
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• 제36권9호
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• pp.1350-1356
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• 2023

Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.