• BMB Reports
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• 제34권6호
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• pp.502-508
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• 2001

A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

• Journal of Animal Science and Technology
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• 제44권6호
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• pp.693-700
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• 2002

한우 종모우에 있어 소 성장호르몬 유전자의 출현빈도를 알아보고 소 성장호르몬 다형과 정액성상과의 관계를 살펴보기 위하여 실시하였다. 한우 종모우 109두의 소 성장호르몬 유전자를 Alu I 제한효소로 처리하여 PCR-RFLP로 분석하였다. Leucine(Leu)과 valine(Val) 유전자의 출현빈도는 각각 0.88과 0.12이었다. 소 성장호르몬 VV 유전자형을 지닌 한우 종모우가 다른 유전자형을 지닌 종모우보다 정액성상(정액량, 정자농도, 총정자수)이 떨어지는 경향을 보였지만 소 성장호르몬의 유전자형이 정액성상에 유의적인 영향은 미치지 못하였다. 채취순번에 따른 영향에 있어서도 VV 유전자형을 지닌 종모우가 다른 유전자형을 지닌 종모우보다 정액성상이 떨어지는 경향을 보였지만 유의적인 차이는 나타내지는 못하였으며 다만 년도에 따른 영향에 있어 1998년도 VV 유전자형을 지닌 종모우가 다른 유전자형을 지닌 종모우보다 총정자수에 있어 유의적으로 적게 나타났다(P<0.05). 전체적으로 소 성장호르몬 유전자의 VV 유전자형을 지닌 종모우의 정액성상이 다른 유전자를 지닌 종모우의 정액성상보다 낮은 경향을 보였지만, 조사한 109두의 한우 종모우 중 VV 유전자형을 지닌 종모우가 1두로 분석되어 이 종모우의 정액성상만을 이용하여 소 성장호르몬 유전자다형이 정액성상에 미치는 영향을 살펴보았기에 종모우 선발시 기초자료로 이용하기에는 추가적인 시험이 필요할 것으로 사료된다.

• Fibers and Polymers
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• 제2권3호
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• pp.141-147
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• 2001

The effect of compositions of isophorone diisocyanate (IPDl)/4.4'-diphenylmethane diisocyalate (MDI) and polypropylene oxide diol (PPG, $M_w$: 3000)/1,4-butane diol (BD) on the properties of UV-cured polyurethane acrylate films based on 2-hydroxyethyl acrylate (HEA) was examined. UV-curable polyurethane acrylates were formulated from the prepolymer. trimethylol propane triacrylate (TMPTA) as a reactive diluent, and 1-hydroxycyclohexyl ketone (Irgacure 184) as a photoinitiator. Dynamic mechanical thermal properties and elastic properties of UV-cured polyurethane acrylates was fecund to depend on the chemical composition of IPDl/MDl and PPG/BD. As the BD content increased, the tensile storage modulus of all series samples increased significantly. The storage modulus increased in the order of samples A (IPDI based samples)> samples B (IPDI/MDl (7/3 molar ratio) based samples) > samples C (IPDI/MDl (5/5 molar ratio) based samples at the same composition. Two distinct louts modulus peaks for all samples are observed owing to the softs segment glass transition temperature ($T_gh$) and hard segment glass transition temperature ($T_gh$). The difference between $T_gh$, and $T_gh$, (Δ$T_g$) increases in the order of A > B > C at the same composition. In cycle test, the initial onset strain (%) was found to decrease with increasing BD content in PPG/BD and with increasing MDI content in IPDI/MDl.

• Animal Bioscience
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• 제34권5호
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• pp.867-879
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• 2021

Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.

• Asian-Australasian Journal of Animal Sciences
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• 제10권4호
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• pp.335-356
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• 1997

Injection of bovine growth hormone (bGH) to lactating dairy cows increases milk yield and yields of milk components including fat. It is generally believed that most of the anabolic effects derived from bGH in animal tissues are primarily mediated by IGF-1. IGF-1 is a strong anabolic peptide in the plasma of animals and exerts mitogenic and metabolic effects on target cells. Contrary to most protein hormones, the majority of IGF-1 in circulation is bound to the binding proteins (IGFBPs) which are known to be responsible for modifying the biological actions of IGF-1, thus making determinations of IGF-1 actions more difficult. On the other hand, fat is a major milk component and the greatest energy source in milk. Currently, the fat content of milk is one of the major criteria used in determining milk prices. It has been known that flavor and texture of dairy products are mainly affected by milk fat and its composition. Acetyl-CoA carboxylase (ACC) is the rate limiting enzyme which catalyzes the conversion of acetyl-CoA to malonyl-CoA for fatty acid synthesis in 1ipogenic tissues of animals including bovine lactating mammary glands. In addition to the short-tenn hormonal regulation of ACC by changes in the catalytic efficiency per enzyme molecule brought about by phosphorylation and dephosphorylation of the enzyme, the long-term hormonal regulation of ACC by changes in the number of enzyme molecules plays an essential role in control of ACC and lipogenesis. Insulin, at supraphysiological concentrations, binds to IGF-1 receptors, thereby mimicking the biological effects of IGF-1. The receptors for insulin and IGF-1 share structural and functional homology. Furthermore, epidermal growth factor increased ACC activity in rat hepatocytes and adipocytes. Therefore, it can be assumed that IGF-1 mediating bGH action may increase milk fat production by stimulation ACC with phosphorylation (short term) and/or increasing amounts of the enzyme proteins (long term). Consequently, the main purpose of this paper is to give the readers not only the galactopoietic effects of bGH, but also the insight of bGH action with regard to stimulating milk fat synthesis from the whole body to the molecular levels.

• Journal of Nutrition and Health
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• 제50권3호
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• pp.236-245
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• 2017

본 연구에서는 난소절제 흰쥐에 구아바잎 추출물을 8주 동안 경구투여한 결과 난소절제 대조군인 OVX군과 비교시 체중 증가량 및 혈중 유리지방산이 유의적으로 감소된 것을 확인하였다. 또한 간 내 중성지방 농도가 $OVX{\cdot}GL$군과 $OVX{\cdot}GH$군에서 모두 유의적으로 감소하였으며 혈중 항산화 효소인 GPx 농도가 유의적으로 증가하였다. 간 내 항산화 효소 및 eNOS의 mRNA 발현 정도를 측정한 결과 OVX군에 비해 구아바잎 추출물 급여군인 $OVX{\cdot}GL$군과 $OVX{\cdot}GH$군에서 모두 Nrf2 및 CAT의 mRNA 발현 정도가 유의적으로 증가하였으며 eNOS또한 $OVX{\cdot}GH$군에서 유의적으로 증가함을 확인할 수 있었다. 따라서 본 연구의 결과를 종합해 볼 때 구아바잎 추출물 경구투여는 항산화 효소의 활성을 증가시키고 혈관내피세포의 기능을 향상시킴으로써 폐경 후 나타날 수 있는 혈관질환과 산화스트레스로 인한 대사적 장애 개선에 도움이 될 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.161-164
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• 2003

The polymorphism of the growth hormone gene in 12 pig breeds (total n=475) was detected by PCR-Apa I-RFLP, and allele A (449 bp, 101 bp and 55 bp) or allele B (316 bp, 133 bp, 101 bp and 55 bp) were observed. In these pig breeds, we found that European pig breeds had high frequencies of allele B, while Chinese native pig breeds had high frequencies of allele A. In addition, the role of porcine GH was investigated in 117 Nanchang White pigs and 361 Large Yorkshire pigs. Eight traits about growth and carcass were recorded for analyzing associations between GH gene polymorphism and performance quantitative traits. In the Nanchang White pigs, no significant difference was observed between different genotypes and different growth and carcass traits. In Large Yorkshire pigs, those with BB genotype had more lean percentage than pigs with AA genotype (p<0.05). Based on these results, we conclude that the GH locus should be further investigated in commercial breeds to determine its suitability for use in marker-assisted selection programmes.

• 한국통신학회논문지
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• 제25권10A호
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• pp.1560-1565
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• 2000

Over an additive abelian group G of order g and for a given positive integer λ, a generalized Hadamard matrix GF(g,λ) is defined as a gλ$\times$gλ matrix [h(i,j)] where 1$\leq$i$\leq$gλ,1$\leq$j$\leq$gλ, such that every element of G appears exactly λ times in the list h(i$_1$,1)-h(i$_2$,1), h(i$_1$,2)-h(i$_2$,2),...,h(i$_1$,gλ)-h(i$_2$, gλ) for any i$\neq$j. In this paper, we propose a new method of expanding a GH(\ulcorner,λ$_1$) = B = \ulcorner over G by replacing each of its m-tuple \ulcorner with \ulcorner GH(g,λ$_2$) where m=gλ$_2$. We may use \ulcornerλ$_1$(not necessarily all distinct) GH(g,λ$_2$)'s for the substitution and the resulting matrix is defined over the group of order g.

• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1090-1094
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• 2006

Histone acetylation modification is one key mechanism in the regulation of gene activation. In this study, we investigated the global levels of histone H4 acetylation of insulin like growth factor I (IGF1) and growth hormone (GH) genes in the lungs of two somatic cell cloned calves. Data showed the levels of histone H4 acetylation of IGF1 and GH genes vary widely within different gene regions, and, in almost all regions of the two genes, acetylation levels are lower in the aberrant clone than in the normal clone. Thus we suggest that inefficient epigenetic reprogramming in the clone may affect the balance between acetylation and deacetylation, which will affect normal growth and development. These findings will also have implications for improvement of cloning success rates.

• 한국유기농업학회지
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• 제27권1호
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• pp.65-76
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• 2019

작물생육촉진과 병 방제 기능을 지닌 Bacillus velezensis GH1-13 균주의 대량배양을 위한 최적배지(glucose 0.5%, soy bean flour 0.8%, NaCl 0.15%, $K_2HPO_4$ 0.25%, $Na_2CO_3$ 0.05%, $MgSO_4.7H_2$ 0.1%) 조성을 확립하였다. 최적배지(MMS)를 이용하여 500 L 대용량 발효기에서 배양한 결과 총 균체수 $7.5{\times}10^9cells/mL$, $6.8{\times}10^9\;endospore\;cells/mL$ 및 90% 내생포자 형성률 등 안정된 대량생산을 확인하였다. 최적배지에서 배양한 GH1-13 균체와 배양 상층액의 경우 Colletotrichum gloeosporioides를 포함한 4종의 식물병원성 곰팡이에 대한 항진균활성을 보였다. 또한 식물생육촉진 호르몬의 일종인 IAA 생산량을 비교한 결과, 최적배지에서 배양한 경우 상업용 배지(TSB, R2A)에 비해 2.5~13배 이상 높은 생산성을 보였다. 더불어, 최적배지에 0.3% tryptophan을 첨가하여 배양했을 경우 28.50 mg/L의 IAA 최대 생산량을 보였으며, 이는 tryptophan을 첨가하지 않고 배양한 경우보다 약 4배 높은 수준이었다. 이러한 결과로 볼 때 본 연구에 사용된 B. velezensis GH1-13 균주는 작물생육촉진 및 곰팡이 병 방제 측면의 복합기능 생물학적 제제로서 매우 유용할 것으로 판단된다.