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닭의 고밀도 사양체계가 스트레스 및 지방대사 연관 유전자 발현에 미치는 영향
안영숙(Young Sook An),박정근(Jeong Geun Park),장인석(In Surk Jang),손시환(Sea Hwan Sohn),문양수(Yang Soo Moon) 한국생명과학회 2012 생명과학회지 Vol.22 No.12
본 연구는 육계에서 고밀도 사양체계가 간의 지놈 전사체, 특히 스트레스 및 지방대사 연관 유전자들의 발현에 어떤 영향을 미치는지 알아보기 위하여 실시하였다. 공시된 시험동물의 대조군 사육밀도는 495 cm2/수, 고밀도군은 245 cm2/수를 35일령까지 유지하였다. 대조구와 비교하여 고밀도 사양 육계에서 체중, 증체량, 사료섭취량이 유의적(p<0.05)으로 감소되는 것으로 나타났다. 폐사율은 고밀도군에서 15.7%로서 대조군(3.7%)에 비해 폐사율이 4.2배 높았다. 육계의 사육밀도에 따른 스트레스관련 유전자 HMGCR, HSP90α, HSPA5 (GRP78/Bip), DNAJC3, ATF4 등의 발현이 증가하였으며, interferon-γ, PDCD4 등의 발현은 감소하였다. Endoplasmic reticulum (ER) stress 관련 HSPA5 (GRP78/Bip), DNAJC3 그리고 ATF4은 유전자들은 고밀도 사양계에서 유전자의 발현이 2-3배 증가함을 보였다. 고밀도 사양은 지방산 합성에 관여하는 효소들(ACSL5, TMEM195, ELOVL6)의 유전자 발현 증가와 지방산산화(β-oxidatin)에 관여하는 효소들(ACAA1, ACOX1, EHHADH, LOC423347, CPT1A)의 RNA 발현 증가를 유도하였다. 본 연구는 밀사에 의한 스트레스가 닭의 간에서 지방을 합성하기 위한 유전자들의 발현을 증가시키고, 합성된 지방산을 분해하여 에너지를 생산하기 위한 지방산의 산화도 높게 유지하고 있음을 보여 주었다. 닭의 주요 지방대사기관인 간에서 외부적 환경인자(사육환경)에 의한 스트레스와 생리적 대사(지방대사 및 소포체 스트레스)가 서로 밀접한 관계가 있음을 분자생물학적 수준에서 확인하였다. 따라서 스트레스저감 사 육환경제공 및 친환경 사육방법 도입 등 동물복지를 고려한 가금사양체계가 필요한 것으로 사료된다. The effect of high stocking density (HSD) on the expression of stress and lipid metabolism associated genes in the liver of broiler chickens was examined by chicken genome array analysis. The chickens in a control group were randomly assigned to a 495 cm2/bird stocking density, whereas the chickens in a HSD group were arranged in a 245 cm2/bird stocking density with feeding ad libitum for 35days. The chickens assigned to the HSD group had a significantly lower body weight, weight gain, and feed intake compared with those of the control group (p<0.05). The mortality of chickens was higher in the HSD group than in the control group. The microarray analysis indicated up-regulation of stress associated genes such as HMGCR, HSP90α, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, and down-regulation of interferon-γ and PDCD4 genes. The endoplasmic reticulum stress associated genes, HSPA5 (GRP78/Bip), DNAJC3 and ATF4, were highly expressed in the HSD group. The genes, ACSL5, TMEM195 and ELOVL6, involved in fatty acid synthesis, were elevated in the HSD group. The genes, ACAA1, ACOX1, EHHADH, LOC423347 and CPT1A, related to fatty acid oxidation, were also activated in the HSD group. These results suggest that a HSD rearing system stimulates the genes associated with fatty acid synthesis as well as fatty acid oxidation in the liver of broiler chickens.
닭의 고밀도사양 스트레스가 품종 간 체내대사 유전자 발현에 미치는 영향
손시환(Sea Hwan Sohn),장인석(In Surk Jang),안영숙(Young Sook An),문양수(Yang Soo Moon) 한국가금학회 2015 韓國家禽學會誌 Vol.42 No.1
& & 본 연구는 밀사에 의한 환경스트레스가 닭의 품종에 따라 스트레스 및 대사 연관 유전자들의 발현에 어떤 영향을 미치는지 알아보고자 실시하였다. 공시계는 한국재래닭과 백색 레그혼으로 두 품종 모두 40주령 때 대조구(540 cm2/수) 및 고밀도구(311 cm2/수)로 분리하고, 50주령까지 10주간 사육하였다. 사양시험 종료 후, 각 개체의 간으로부터 total RNA를 추출하고, 스트레스, 소포체(ER) 스트레스 및 대사 연관 유전자들의 발현을 real-time PCR을 이용하여 분석하였다. 한국재래계는 분석된 모든 스트레스 표지 유전자들의 발현이 밀사구와 대조구 사이에 유의적인 변화를 보이지 않았다. 그러나 백색레그혼의 경우, HSP70과 HSP90α 유전자의 발현이 유의적으로 높게 나타났다(P<0.05). 분석된 ATF6, GRP78, SREBP2 등의 발현은 품종 간 차이를 볼 수 없었지만, XBP1의 경우 백색레그혼이 한국재래계에 비하여 높은 발현을 보였다(P<0.05). 분석된 유전자들 중 FABP4, FATP1, ACSL1 등의 경우, 한국재래계에 비하여 백색레그혼에서 높은 유전자 발현을 보였다(P<0.05). GLUT의 발현은 품종 간에는 영향을 받지 않지만, 밀사에 의한 영향을 받고 있음을 보여주었다. 고밀도사양 체계는 닭의 품종과 관계없이 스트레스 요인이 될 수 있으며, 닭의 품종이나 개량의 정도에 따라 스트레스 반응에 대한 유전적 차이가 있음을 시사하고, 또한 밀사와 같은 환경적 스트레스는 간의 지방 및 포도당 대사에 영향을 미칠 수 있음을 보여주었다. & & Chickens are exposed to the external and internal stressors such as low and high temperature, high stocking density, feed restriction and disease. There have been a few studies on gene expressions through the investigation of chickens under direct exposure to the stress of high stocking density. The objective of the present study was to determine the expressions of genes associated with stress, endoplasmic reticulum (ER)-stress, lipid and glucose metabolism in two strains of chickens, Korean Native Chicken (KNC) and White Leghorn (WL), raised in high stocking density. A total of 164 chickens aged 40 weeks were randomly allotted to a 540 cm<SUP>2</SUP>/bird stocking density (control), whereas the chickens in a high density group were assigned in a 311 cm<SUP>2</SUP>/bird stocking density with feeding ad libitum for 10 weeks. Total RNA was extracted from the live for qRT-PCR. The expression levels of hsp70 and hsp90α were higher in WL subjected to stress with high stocking density compared with those genes in control (P<0.05), while the expressions of genes were not affected in KNC. ER stress marker gene XBP1 was also highly expressed in WL with stress (P<0.05), but the stress of high stocking density did not influence to ER stress marker genes in KNC. Lipid metabolism associated genes including FABP4, FATP1 and ACSL1 were highly expressed in WL compared with KNC when subjected to high stocking density stress (P<0.05). The expression of glucose transport gene GLUT2 and GLUT8 were increased in chickens exposured to the stress of high stocking density (P<0.05). The data indicate that WL is more sensitive to the stress of high stocking density compared with KNC and the stress may influence the modulation of lipid and glucose metabolism in the liver of chickens.
Helicobacter pylori와 대장균의 Shuttle Vector 개발
조명제,이우곤,이상룡,김경희,안영숙,김성희,김현주,류복덕,최여정,윤영혜,백승철,전영석,이광호 경상대학교 유전공학연구소 1997 遺傳工學硏究所報 Vol.16 No.-
In this study, a vehicle vector using cryptic plasmids was constructed for gene transfer in Helicobacter pylori. pHP51(3.9 kb) and pHP489(1.2 kb) were selected for constructing vectors from cryptic plasmid of H. pylori isolates in Korea. The HindⅢ-digestedDNA fragment(1.2kb) of pHP489 and 1.6kb DNA fragment of pHP51 were ligated with a kanamycin resistance gene(aph3'-Ⅲ) from C. jejuni to produce the recombinant plasmids pHP489K and pHP51K, respectively. Transformation frequency of pHP51K by electroporation was low. But pHP489K could be effectively transformed into various H. pylori strains. In order to design an intermdiate vehicle vector for gene transfer into H. pylori, pBlueHP489K was prepared by recloning pHP489K DNA into pBluescript and pTZ19R vector. This vector permitted the DNA fragment containing pHP489 sequence, aph3'-Ⅲ, and cloned DNA to be cut and self-ligated in the SacⅠ site after cloning. ureA and ureB gene were inserted into pBlueHP489K, resulting in pBlueHP489K/AB. The DNA fragment containing pHP489, kanamycin resistance gene(aph3'-Ⅲ), and urease structural gene was cut away from pBlueHP489K/AB and self-ligated to generate pBlueHP489K/AB. pBlueHP489K/AB made urease-negative H. pylori strains restore their urease activity. By this experiment, pBlueHP489K was confirmed to be the vehicle system for transferring H. pylori genes.
원료형태와 추출 용매, 시간 및 온도 조건에 따른 미강으로부터의 Tocopherol 및 Tocotrienol 추출 효율 비교
안영숙,이영상 순천향대학교 기초과학연구소 2004 순천향자연과학연구 논문집 Vol.10 No.2
The rice bran contains various nutraceutical compounds such as vitamin E consisting of 8 isomers of tocopherols and tocotrienols. Aiming at industrial utilization of rice bran as a supply for vitamin E, various conditions to extract tocopherols and tocotrienols from fresh bran powder, bran pellet, and crude oil (hexane extracts of bran) were tested and the efficiency of extraction was compared. Hexane proved the most suitable extracting solvent when safety and economic concerns were considered although the relative efficiency of recovery was petroleum ether (1.3)>ethyl ether (1.2)>acetone (1.1)>hexane(1.0)>heptane (1.0)> isopropyl alcohol(0.9). Extracting bran pellet at room temperature exhibited similar vitamin E recovery compared to extraction at 60℃, suggesting no necessity of additional heating during extraction. Based upon higher recovery rate and convenience of handling, hexane-extraced crude oil, corresponding to 15% weight of bran powder, was the most pertinent form of bran to extract tocopherols and tocotrienols compared to fresh bran powder, was the most pertinent form of bran to extract tocopherols and tocotrienols compared to fresh bran powder and pellet.
미강 함유 Tocopherol 및 Tocotrienol 추출을 위한 최적 Saponification 조건의 탐색
안영숙,이영상 순천향대학교 기초과학연구소 2004 순천향자연과학연구 논문집 Vol.10 No.2
Tocotrienols and tocopherols rich in rice bran are functional compounds collectively called as vitamin E bearing antioxidant, anticancer, and cholesterol-lowering effects. Various factors determining the recovery of tocopherol and tocotienols during the saponification to extract these compounds from rice bran crude oil were tested. Regarding the ratio of crude oil weight to EtOH volume for saponification, up to 15 g crude oil in 50 mL EtOH resulted in relatively high recovery of total vitamin E 1.3 mg·g^(-1) crude oil, which was only 3 % loss compared to 5 g crude oil. In case of 5 g crude oil was to be saponified in 50 mL EtOH, the optimum amount of 44 % KOH was 2 mL and the recovery rate under such condition was total vitamin E 1.2mg·g^(-1) crude oil. However, insufficient saponification caused by adding higher amount of crude oil and less amount of KOH produced undesirable residues even after evaporating hexane layers containing unsaponifiable fractions, which in turn resulted in lower concentration of tocopherols and tocotrienols in final products. Re-saponification of such residues significantly decreased recovery by 30% to 60%. Response-surface analysis to determine optimal temperature and duration for saponification indicated 83.7℃ and 18 min when 5 g crude oil was added into 50 mL EtOH and 2 mL KOH was used.