• 제목/요약/키워드: MS-222

검색결과 63건 처리시간 0.03초

CCoAOMT 유전자 발현 억제에 의한 현사시나무의 화학조성 변화 (Characterization of Chemical Composition in Poplar wood (Populus deltoides) by Suppression of CCoAOMT Gene Expression)

  • 엄인용;김광호;이수민;이용섭;최준원
    • Journal of the Korean Wood Science and Technology
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    • 제38권3호
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    • pp.213-222
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    • 2010
  • 본 연구에서는 리그닌 전구물질 생합성에 관여하는 CCoAOMT (Caffeoyl-CoA-O-methyltransferase)유전자의 발현을 분자생물학적으로 억제시킨 상태와 정상적인 상태에서 생장한 현사시나무를 대상으로 홀로셀룰로 오스, 리그닌과 단당류 조성에 관한 정량 분석을 각각 실시하였다. 그리고 각각의 목질바이오매스로부터 MWL (milled wood lignin)을 추출하여 메톡실기정량, DFRC, Py-GC/MS, GPC, $^{13}C$ NMR 분석 등 리그닌의 화학 구조적 특성을 비교하였다. 정상재와 형질전환체의 홀로셀룰로오스 함량은 각각 81.6%와 82.3%로 큰 차이는 없었지만, 리그닌 함량은 각각 21.7%와 18.3%로 형질전환체가 약 3.4% 정도 낮았다. 정상재와 형질전환체의 구성 단당류 분석결과, 글루코오스는 각각 511.0 mg/g와 584.8 mg/g, 그리고 자일로오스는 각각 217.8 mg/g와 187.5 mg/g로 나타나 CCoAOMT 발현억제는 단당류 조성에 영향을 미치는 것으로 분석되었다. 각 시 료로부터 단리한 리그닌(MWL)의 메톡실기 정량에 따르면 CCoAOMT 발현을 억제한 시료에서 약 7% 정도의 메톡실기가 감소하는 것으로 나타났고, 이는 $^{13}C$-NMR 분석과 Py-GC/MS 분석을 통하여 확인할 수 있었다. 또한 Py-GC/MS 분석에 의하면 정상재와 형질전환체 리그닌의 G/S 비율이 각각 0.59와 0.44로 나타난점으로 미루어 CCoAOMT 발현 억제는 리그닌 생합성 과정에서 G unit 생합성보다 S unit 생합성에 더 많은 영향을 미치는 것으로 예측할 수 있다.

Siberian Wildrye Grass의 성숙종자 유래의 캘러스로부터 식물체 재분화 (In Vitro Plant Regeneration of Siberian Wildrye Grass from Mature Seed-derived Callus)

  • 이기원;오치르바트 친저르익;최기준;윤세형;김기용;지희정;이상훈
    • 한국초지조사료학회지
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    • 제31권3호
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    • pp.217-222
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    • 2011
  • Siberian wildrye grass는 몽골지역 방목 초지에 넓게 분포하는 대표적인 초종으로 사료가치가 높고 장기간 이용 할 수 있는 장점 등으로 그 중요성이 증대되고 있다. 그러나 최근 기후 변화로 인한 사막화로 초지의 면적이 점점 줄어들고 있는 추세이다. 따라서 환경에 대한 적응성이 우수한 신품종을 개발할 목적으로 우선 효율적인 기내배양 조건을 확립하고자 하였다. Siberian wildrye grass의 최적의 기내배양 조건을 확립하기 위하여 몽골에서 수집한 성숙종자로부터 최적 캘러스 유도조건 및 효율적인 식물체 재분화 체계를 확립하였다. 캘러스는 5mg/L 2,4-D가 첨가된 MS 배지에서 가장 높은 빈도로 유도되었으며, 식물체 재분화는 1 mg/L 2,4-D와 3 mg/L BA가 첨가된 N6 배지에서 캘러스를 배양했을 때 50% 이상의 재분화율을 나타내었다. 본 연구를 통하여 확립된 기내배양 시스템은 분자육종을 통한 신품종 siberian wildrye grass의 개발에 유용하게 이용되어질 수 있을 것이다.

Identification of Fatty Acids in the Oils of Pine Nuts by GC-MS of Their Picolinyl Esters and 4,4-dimethyloxazoline Derivatives in Combination with Silver-Ion Chromatography

  • Kim, Seong-Jin;Woo, Hyo-Kyeng;Seo, Min-Young;Joh, Yong-Goe
    • 한국응용과학기술학회지
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    • 제19권3호
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    • pp.222-244
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    • 2002
  • A mixture of methyl ester derivatives of fatty acids from the oils of pine nuts was well resolved to five fractions differing by degree of unsaturation by silver ion solid-phase extraction column chromatography ($Ag^{+}$-SEC). Polyunsaturated fatty acid with non-methylene interrupted conjugated double bond (NMiDB) radical held more strongly to silver ions in the column than methylene interrupted conjugated double bond (MiDB) one when they had the same number of double bonds. Although both the picolinyl ester and DMOX derivative provided clear mass ion species powerful enough to elucidate the structure of the polyunsaturated fatty acid (PUFA) with NMiDB and/or methylene interrupted conjugated double bond (MiDB) radical in the oils, the picolinyl ester of PUFA with NMiDB radical did not provide a cluster of mass ions neighboring diagnostic mass ions induced by the double bond in the proximal to the carboxyl group. However, the DMOX derivative of PUFA with NMiDB group as well as MiDB showed abundant mass ion species differing by gaps of 12 amu, which made it possible with greater ease to locate the double bonds in the molecule. The oil contained $C_{18:2{\omega}6}$ (46.2 %) and $C_{18:1{\omega}9}$ (25.4 %) as main components, and considerable amounts of PUFAs with NMiDB radical such as ${\Delta}^{5.\;9.\;12}-C_{18:3}$ (16.0 %), ${\Delta}^{5.\;9}-C_{18:2}$ (2.3 %) and ${\Delta}^{5.\;11.\;14}-C_{20:3}$ (0.8 %).

Altered Proteome of Extracellular Vesicles Derived from Bladder Cancer Patients Urine

  • Lee, Jingyun;McKinney, Kimberly Q.;Pavlopoulos, Antonis J.;Niu, Meng;Kang, Jung Won;Oh, Jae Won;Kim, Kwang Pyo;Hwang, Sunil
    • Molecules and Cells
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    • 제41권3호
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    • pp.179-187
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    • 2018
  • Proteomic analysis of extracellular vesicles (EVs) from biological fluid is a powerful approach to discover potential biomarkers for human diseases including cancers, as EV secreted to biological fluids are originated from the affected tissue. In order to investigate significant molecules related to the pathogenesis of bladder cancer, EVs were isolated from patient urine which was analyzed by mass spectrometry based proteomics. Comparison of the EV proteome to the whole urine proteome demonstrated an increased number of protein identification in EV. Comparative MS analyses of urinary EV from control subjects and bladder cancer patients identified a total of 1,222 proteins. Statistical analyses provided 56 proteins significantly increased in bladder cancer urine, including proteins for which expression levels varied by cancer stage (P-value < 0.05). While urine represents a valuable, non-invasive specimen for biomarker discovery in urologic cancers, there is a high degree of intra- and inter-individual variability in urine samples. The enrichment of urinary EV demonstrated its capability and applicability of providing a focused identification of biologically relevant proteins in urological diseases.

Gas Exchange Rates Measured Using a Dual-Tracer ($SF_6$ and $^3He$) Method in the Coastal Waters of Korea

  • Lee, Hyun-Woo;Lee, Ki-Tack;Kaown, Duk-In
    • Ocean Science Journal
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    • 제43권1호
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    • pp.17-24
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    • 2008
  • Over a period of 5 days between August 12 and 17, 2005, we performed a gas exchange experiment using the dual tracer method in a tidal coastal ocean located off the southern coast of Korea. The gas exchange rate was determined from temporal changes in the ratio of $^3He$ to $SF_6$ measured daily in the surface mixed layer. The measured gas exchange rate($k_{CO_2}$), normalized to a Schmidt number of 600 for $CO_2$ in fresh water at $20^{\circ}C$, was approximately $5.0\;cm\;h^{-1}$ at a mean wind speed of $3.9\;ms^{-1}$ during the study period. This value is significantly less than those obtained from floating chamber-based experiments performed previously in estuarine environments, but is similar in magnitude to values obtained using the dual tracer method in river and tidal coastal waters and values predicted on the basis of the relationship between the gas exchange rate and wind speed (Wanninkhof 1992), which is generally applicable to the open ocean. Our result is also consistent with the relationship of Raymond and Cole (2001), which was derived from experiments carried out in estuarine environments using $^{222}Rn$ and chlorofluorocarbons along with measurements undertaken in the Hudson River, Canada, using $SF_6$ and $^3He$. Our results indicate that tidal action in a microtidal region did not discernibly enhance the measured $k_{CO_2}$ value.

난황형성부터 성숙까지 메기(Silurus asotus) 난모세포의 미세구조 (Ultrastructural Changes of Oocytes from Vitellogenesis into Maturation in Korean Catfish, Silurus asotus)

  • 윤종만;김영길
    • Applied Microscopy
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    • 제28권4호
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    • pp.447-463
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    • 1998
  • This study was carried out to investigate the histomorphological changes and the electrophoretic patterns of egg components, obtained from 100 of 2-year-old female catfishes (Silurus asotus). Female Korean catfishes collected in the vicinity of Chollabukdo have synchronous ovaries which discontinously ovulate eggs during the breeding season (from late May until early July). The fishes were killed by severing the spinal cord just posterior to the head after immobilization with tricaine methanesulfonate (MS 222). Especially, the light microscopic and ultrastructural changes of ooplasm and follicular membranes of oocytes, were examined by means of light and transmission electron microscopy. The size of the nucleoli and number of the yolk granules increased as the oocyte groved. Yolk granules were deposited in the oocyte as fluid Due to tile presence of large early and late maturing oocytes, their ovaries were enlarged, transparent, granular and greenish in color. As the percentages of fish in late maturing oocyte (LMO) and ripe oocyte (RO) stage increased from March to April, mean gonadosomatic index (GSI) values (19.95%) increased. Zona radiata changed from squamous into cuboid in shape in the early maturing oocyte (EMO) stage. Processes, microvilli, from the zona radiata and from the oocyte grow, and make contact with each other in the pore canals of the zona radiata during vitellogenesis, but are withdrawn as the zona radials becomes more compact and devoid of pore canals during oocyte maturation. Seasonal changes in the microscopic appearance of the ovaries were well correlated with those in both gonadosomatic index and macroscopic appearance. The main cytological changes such as increase in size of cell, nucleus, nucleolus, and increase in number of nucleoli and mitochondria demonstrated with electron microscopy in the previtellogenic oocytes of Korean catfish, provided evidences for important synthetic processes in an early preparatory phase of oocyte development. The electrophoretic pattern of major band in mature stage was much thicker (24 k, 66 k, 90-110k dalton) than that in previtellogenic phase.

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유통 환(丸)제품의 중금속 함량 및 위해성 평가 (The Content of Heavy Metals in Herbal Pills Used as General Processed Food and Risk Assessment of Heavy Metal Intakes)

  • 김성단;정선옥;김복순;윤은선;장민수;박영애;이용철;채영주;김민영
    • 한국식품영양과학회지
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    • 제39권7호
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    • pp.1038-1048
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    • 2010
  • 본 연구는 시중에 유통되고 있는 환(丸)형태 일반가공식품 18종 총 52건을 microwave로 습식분해 후 ICP-MS 및 Mercury Analyzer를 이용하여 납, 카드뮴, 크롬, 구리, 수은 잔류량을 분석한 결과, 평균 0.421(0.032~1.630), 0.157(0.011~0.515), 1.033(0.112~9.933), 6.923(1.333~16.755), 0.010(0.001~0.088) mg/kg이 검출되었다. 사용된 부위별 납 검출 수준을 살펴보면 잎(Herba, 1.032 mg/kg)이 다른 부위에 비해서 유의적인 수준으로 높았으며, 열매(fructus, 0.222 mg/kg), 뿌리줄기(Rhizoma, 0.179), 씨(semen, 0.153 mg/kg) 부위의 납 함량이 유의적으로 낮았다(p<0.05). 카드뮴 또한 해조류(algae, 0.304 mg/kg), 잎(0.298 mg/kg)을 사용한 제품에서 유의적인 수준으로 높게 잔류하였다(p<0.05). 또한 카드뮴은 다시마를 원료로 한 환제품 총 4건 중 2건(0.431, 0.370 mg/kg)에서, 인진쑥환 총 7건 중 5건(0.315~0.515 mg/kg)에서, 당귀환 1건(0.338 mg/kg), 표고버섯환 1건(0.394 mg/kg)에서 한약재 기준인 0.3 mg/kg을 초과하는 수준으로 검출되었다. 따라서 환(丸)형태 일반가공식품에 대한 카드뮴의 조속한 기준설정 및 관리가 필요한 실정이다. 대부분의 중금속 잔류량이 높았던 잎(Herba)의 중금속별 평균 함량은 Cu 6.480 mg/kg> Cr 2.291 mg/kg> Pb 1.032 mg/kg> Cd 0.298 mg/kg> Hg 0.016 mg/kg 순서로 나타났다. 환(丸)형태 일반가공식품 섭취 시 중금속에 대한 안전성 평가를 위해 제품을 통한 중금속의 주간섭취량과 FAO/WHO에서 제시하는 잠정주간섭취허용량(PTWI), 1인 1일 최대섭취허용량(PMTDI)과 미국환경보호청(U.S. EPA)의 만성경구섭취 참고용량(RfD)과 비교하여 %PTWI를 구하였다. 금속별 주간 섭취량과 %PTWI는 납, 카드뮴, 크롬, 구리, 수은 각각 평균 0.280(0.018~1.272), 0.113(0.002~0.648), 0.630(0.053~4.975), 6.366(0.289~43.217), 0.011(0.000~0.210) ${\mu}g/kg$ bw/week이었으며, %PTWI는 1.112(0.072~5.088), 1.614(0.029~9.257), 3.000(0.252~23.690), 0.182(0.008~1.235), 0.220(0.000~0.420)%로 환(丸)형태 일반가공식품을 통한 중금속 섭취량은 안전한 수준으로 판단되었다.

활어수송시 수온, 염분 및 마취제에 의한 넙치(Paralichthys olivaceus)의 스트레스 반응

  • 허준욱;민병화;이복규;박인석;장영진
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2003년도 추계학술발표대회 논문요약집
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    • pp.103-104
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    • 2003
  • 어류의 양식생산 과정중에는 사육중인 어류에게 스트레스로 작용할 수 있는 여러 가지 요인들이 있다. 그 중에서도 생산된 종묘의 양식장 운송, 수확한 어류의 소비지 운반 등은 양식과정중 피할 수 없는 작업이다. 어류의 수송 후에는 혈장 corticosteroid, glucose, 전해질, 삼투질 농도, 적혈구수, hematocrit, hemoglobin 등이 변화하는 것으로 알려지고 있다(Chang et al. 2001; Hur et al., 2002, 2003). 스트레스 요인별 연구로는 염분, 수온, 밀도 및 마취제에 관한 것 등이며, 아울러 이들의 복합적인 요인에 대하여 스트레스 반응을 최소화시키려는 연구가 진행되고 있다. 어류종묘의 수송이나 수확된 어류가 대량으로 수송되고 있으므로, 넙치(Paralichthys olivaceus)와 같은 대량수송 어류의 수송에 따른 스트레스 반응에 대한 연구의 필요성이 제기된다. 그러므로 본 연구에서는 넙치를 사용하여 염분, 수온 및 마취제에 의한 수송이 스트레스 지표로 알려져 있는 혈액학적 요인, cortisol, glucose, lactic acid 및 삼투질 농도 둥에 나타나는 생리학적 반응을 조사하여, 활어수송 과정에서 나타나는 스트레스 반응에 대한 기초자료를 제공하고자 하였다. 실험어는 21.2 cm, 97.4 g인 양식 넙치를 사용하여, 수온은 20℃ (natural water temperature, NWT)와 15℃ (cooling water temperature, CWT), 염분은 해수(35‰)와 15‰해수, 마취제(anesthesia, Anes., MS-222)는 50 ppm의 조건으로 혼합한 실험구를 설정하였다. 실험구는 각각 NWT+35‰, CWT+35‰, NWT+15‰, NWT+15‰, NWT+35‰+Anes., CWT+35‰+Anes., NWT+15‰+Anes. 및 CWT+15‰+Anes.의 8개 실험구를 2반복으로 설정하여 경북울진∼부산까지 약 400 km (6시간)를 차량수송하였다. 수송용기는 스티로폼상자(66×42×20 cnn)로서, 여기에 해수 3 L와 액화산소를 넣은 비닐봉지에 넙치 8마리씩 수용하여 수송하였다. 혈액의 성상 및 분석항목은 수송전ㆍ후에 채혈하여 비교하였다. 수송전 hematocrit는 22.2±3.8%에서 수송후 NWT+35‰에서 15.3+3.9%, CWT+35‰은 16.7±3.0%, NWT+15‰구에서는 19.2±1.8%로 낮아졌으며, CWT+15‰구는 20.9±3.6%로 수송전과 차이가 없었다. 한편 NWT+15‰+Anes.구는 17.8±0.9%, CWT+15‰+Anes.구는 14.5±1.5%로 낮아졌다. Cortisol은 수송전 2.4±0.1 ng/ml로부터 CWT+35‰구는 16.7±12.8 ng/ml, NWT+35‰구는 47.9+19.8 ng/ml, NWT+15‰구는 43.5±13.9 ng/ml, CWT+15‰구는 26.1±8.3 ng/ml, NWT+15‰+Anes.구는 61.7±3.3 ng/ml, CWT+15‰+Anes.구는 86.1±19.0 ng/ml로 높아졌다. Glucose는 수송전 74.2±32.6 mg/dl로부터 NWT+35‰구는 197.9±27.5 mg/dl, CWT+35‰구도 272.1±29.9 mg/dl로 유의하게 높아졌다. Na/sup +/의 수송전 농도는 163.5±0.6 mEq/L로부터 NWT+35‰구와 CWT+35‰구는 각각 175.3±1.2 mEq/L, 190.0±5.0 mEq/L로 높아졌으며, 다른 실험구에서는 차이가 없었다. 본 연구 결과, cortisol과 glucose에서 수송전보다는 모든 실험구에서 높게 나타나 수온, 염분 및 마취제를 사용하여도 스트레스를 받고 있는 것으로 나타났다. 특히, cortisol의 경우, 수온과 염분만을 혼합한 실험구보다 마취제를 혼합한 실험구에서 높게 나타났다. 다른 혈액학적 항목에서는 염분과 마취제를 사용하지 않았던 실험구인 NWT+35‰구와 CWT+35‰구에서 다른 실험구에 비하여 glucose, Na/sup +/ 및 Cl/sup -/ 등에서는 높게 나타나는 경향을 보였다.

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Solubilization of Proteins from Human Lymph Node Tissue and Two-Dimensional Gel Storage

  • De Marqui, Alessandra Bernadete Trovo;Vidotto, Alessandra;Polachini, Giovana Mussi;De Mattos Bellato, Claudia;Cabral, Hamilton;Leopoldino, Andreia Machado;De Gois Filho, Jose Francisco;Fukuyama, Erica Erina;Settanni, Flavio Aurelio Parente;Cury, Patricia Maluf;Bonilla-Rodriguez, Gustavo Orlando;Palma, Mario Sergio;Tajara, Eloiza Helena
    • BMB Reports
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    • 제39권2호
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    • pp.216-222
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    • 2006
  • In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

Effects of replacing fish oil with palm oil in diets of Nile tilapia (Oreochromis niloticus) on muscle biochemical composition, enzyme activities, and mRNA expression of growth-related genes

  • Ayisi, Christian Larbi;Zhao, Jinliang;Yame, Chen;Apraku, Andrews;Debra, Grace
    • Fisheries and Aquatic Sciences
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    • 제22권11호
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    • pp.25.1-25.9
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    • 2019
  • Background: Due to the continuous demand for fish coupled with decline in capture fisheries, there is the need to increase aquaculture production to meet the demand. Aquaculture is faced with high cost of feeding since fish oil and fish meal are expensive. In view of this, there are calls to explore alternatives that are cheap and reliable. Objectives: This study on Oreochromis niloticus was conducted to evaluate the effects of replacing fish oil (FO) with palm oil (PO) at 0%, 25%, 50%, 75%, and 100% on muscle fatty acid and proximate composition as well as growthrelated enzyme activities and mRNA expression. Methods: Oreochromis niloticus were fed five experimental diets (33% crude protein and 10% crude lipid) for 8 weeks. Feed had variation in fish oil and palm oil contents. After the 8 weeks feeding trial, five fish were sampled from each tank (15 from each treatment) and euthanized using an excess dose of tricaine methane sulfonate (MS-222 at 200 mg/L). Fatty acid and enzyme activities were analyzed using standard protocols. Also, RT-qPCR was used to quantify the expression levels of selected growth-related genes. Results: Fish fed 25% PO recorded the least muscle protein content and was significantly lower than the group fed 100% PO. Paired box protein 7 (Pax-7) enzyme activity was significantly higher in the group fed 50% PO compared to the groups fed 25% PO and 100% PO, while caplain-3 (Capn-3) was significantly lower in the group fed 0% PO compared to all other groups. There was a significant difference among treatments with respect to mRNA expression of Pax-7 and Capn-3. Group fed 25% PO had significantly lower mRNA expression of Pax-7, while the group fed 75% PO recorded significantly higher mRNA expression of Capn-3 compared to groups fed 0% PO, 25% PO, and 100% PO. Pearson's correlation analysis revealed that Igf-I and Igf-II mRNA expression have significant correlation with n-3 polyunsaturated fatty acids content in muscle. Conclusion: The results suggest muscle protein content could be modified if FO is replaced with PO. Also, mRNA expression of Pax-7 and Capn-3 is affected by replacing FO with PO.