해조류 추출물로부터의 Photosensitizing 효과를 지니는 항 적조물질 탐색

진형주 ( Hyung Joo Jin ),진덕희 ( Deuk Hee Jin ) 한국수산과학회 2007 한국수산과학회지 Vol.40 No.3

해조류 김 Porphyra yezoensis 엽체로부터 산에 민감한 유전자의 분리

진형주(Hyung-Joo Jin),박선미(Sun-Mee Park),Long-Guo Kim,진덕희(Deuk-Hee Jin),공인수(In-Soo Kong),홍용기(Yong-Ki Hong) 한국생물공학회 2000 KSBB Journal Vol.14 No.6

개다시마 Kjellmaniella crassifolia Miyabe (Phaeophyta)의 암,수배우체 분리 및 보존

김진희 ( Jin Hee Kim ),김영대 ( Young Dae Kim ),송홍인 ( Hong In Song ),공용근 ( Yong Gun Gong ),이상범 ( Sang Beum Lee ),진형주 ( Hyung Joo Jin ) 한국수산과학회 2007 한국수산과학회지 Vol.40 No.6

Effects of Myostatin Prodomains on the Reproduction of Rotifer Brachionus rotundiformis

Mi Jin Jo(조미진),Hyung-Joo Jin(진형주) 한국생명과학회 2012 생명과학회지 Vol.22 No.4

Myostatin (MSTN)은 TGF (transforming growth factor)-beta family에 속하며, 골격근 성장의 억제 조절인자로서 여러 포유류에서 MSTN 유전자 돌연변이는 골격근 증가를 유도한다. MSTN prodomain은 MSTN의 생물학적 활성을 저해하는데, MSTN prodomain이 과 발현된 쥐에서 과도한 근육축적이 확인되었다. 로티퍼(rotifer;Brachionus rotundiformis)는 치어기 어류의 양식산업에 있어 주요한 일차적 먹이생물이다. 그러나 로티퍼에서 MSTN 및 MSTN prodomain의 기능과 발현 유무는 알려져 있지 않다. 따라서 본 연구는 재조합 MSTN prodomains 이로티퍼에 미치는 영향에 관하여 조사하고자 하였다. 로티퍼 개체배양 실험을 통하여 재조합 MSTN prodomains(pMALc2x-poMSTNpro, pAMLc2x-sMSTNpro)에 의한 로티퍼의 생식 전 단계, 순 생식단계, 생식 후단계, 산란, 수명, 포란, 수컷 발생률을 확인하였으며, 또한 pMALc2x-poMSTNpro와 pAMLc2x-sMSTNpro이 밀집배양에서 로티퍼의 개체성장에 영향을 미치는지에 대하여 확인하였다. 그 결과 농도가 1, 2, 4 μg/ml에서 pMALc2x-poMSTNpro를 처리한 실험군과 0.25 μg/ml에서 4 μg/ml 농도까지 pMALc2x-sMSTNpro를 처리한 실험군에서 로티퍼의 생식 전 단계가 아무처리하지 않은 대조군에 비하여 짧아졌다. 밀집배양 실험에 있어 pMALc2x-poMSTNpro와 pMALc2x-sMSTNpro 모두 로티퍼의 개체 수를 증가를 유도하여, 재조합 MSTN prodomains에 의해서 로티퍼의 reprodution에 영향을 주는 것으로 나타났다. 하지만, 재조합 MSTN prodomains이 어떠한 수용체를 이용하여 신호를 전달하는지에 대한 연구는 앞으로 더 진행되어야 하며, 본 연구의 결과는 재조합 MSTN prodomains이 미세조류에서의 기능 및 메커니즘연구에 중요한 기초자료가 될 것으로 사료된다. Myostatin (MSTN), a member of the transforming growth factor (TGF)-beta family, is a potent negative regulator of skeletal muscle growth and maintenance. The MSTN prodomain inhibits MSTN biological activity. The rotifer Brachionus rotundiformis is an excellent primary live feed for fish larvae in aquaculture; however, it is not known whether the rotifer expresses MSTN and the MSTN prodomain along with its activity. The objective of this study was to examine the effects of recombinant MSTN prodomains. Individual cultures of the rotifer B. rotundiformis were carried out to determine the effect of recombinant MSTN prodomains (pMALc2x-poMSTNpro and pMALc2x-sMSTNpro) on the pre-reproductive phase, reproductive phase, post-reproductive phase, offspring, lifespan, fecundity, and male ratio. In addition, a population culture of the rotifer was performed to confirm the effects of pMALc2x-poMSTNpro and pMALc2x-sMSTNpro on population growth. The results showed that the rotifer treated with pMALc2x-pMSTNpro had a reduced pre-reproductive phase at higher concentrations (1, 2, and 4 μg/ml) compared to the non-treated control group. Moreover, the pMALc2xsMSTNpro treated rotifer effectively decreased the pre-reproductive phase at a lower concentration (0.25 μg/ml) compared to the pMALc2x-pMSTNpro treated and control group. Interestingly, pMALc2x-poMSTNpro and pMALc2x-sMSTNpro significantly increased the population of B. rotundiformis.

유전자증폭반응 기법을 이용한 홍조류 잔금분홍잎 및 누은분홍잎의 구별

김용국(Long Guo Kim),진형주(Hyung Joo Jin),김영식(Young Sik Kim),박중연(Jung Youn Park),남기완(Ki Wan Nam),홍용기(Yong Ki Hong) 한국수산과학회 1997 한국수산과학회지 Vol.30 No.4

The polymerase chain reaction(PCR) technique was used to distinguish from two morphologically similar red algal species; Acrosorium polyneurum and A. yendoi. Total DNA was extracted by the LiCl method. The extracted DNA(15ng) in a 25㎕ reaction volume was amplified by the PCR technique using primers covering with mitochondrial D-loop gene, nuclear rDNA internal transcribed spacer(ITS), and nuclear rDNA external transcribed spacer. A. yendoi could be distinguished from A. polyneurum on the producible basis of amplified ITS fragment of 650bp.

Annealing control primer system을 이용한 어류 재조합 myostatin prodomain 단백질에 의해 성장이 증가된 무지개송어의 특이적 발현 유전자 탐색

이상범 ( Sang Beum Lee ),진형주 ( Hyung Joo Jin ) 한국어류학회 2012 韓國魚類學會誌 Vol.24 No.2

이전 연구에서 넙치유래 재조합 마이오스타틴 프로도메인을 무지개송어에 한달간 침지법을 통하여 처리한 결과 대조군에 비하여 무게가 최대 약 42% 증가되었다. 따라서 본 연구는 재조합 마이오스타틴 프로도 메인을 침지법에 의해 처리된 무지개송어와 대조군의 근육으로부터 발현되는 cDNA를 제작하여 마이오스타틴 프로도메인에 의해서 유도된 특정유전자를 선발하기 위하여 ACP (annealing control primer)를 이용한 DDRT법 을 통하여 분석하였다. 총 20가지의 ACP를 이용한 결과 2개의 특정 유전자를 분석하였으며, NCBI BLAST분석 결과 Cytochrome P450 mono oxygenase와 Profilin으로 판명되었다. 이 중 Cytochrome P450 mono oxygenase는 대조군보다 발현량이 증가하였으며, Profilin는 대조군에 비해서 발현량이 감소하였다. 이러한 결과를 재확인하기 위하여 두 유전자의 primer를 각각 제작하여 semi-quantitative RT-PCR를 시행한 결과 DDRT법에 의한 분석과 동일하였다. 본 결과는 어류의 성장에서 마이오스타틴 프로도메인의 기능 및 메카니즘에 대한 연구에 유용한 자 료가 될 것으로 사료된다. The present study was conducted to investigate different gene expression profile between treated poMSTNpro and non-treated in rainbow trout and to identify those genes that are specifically or predominantly expressed in treated poMSTNpro by employing annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR). We isolated total RNAs in muscle tissues from the treated poMSTNpro fish by immersion bath technique with fish myostatin prodomain (Paralichthys olivaceus, poMSTNpro) for one month and the other was non-treated poMSTNpro, and synthesized cDNA using annealing control primers (ACP, Seegene, Korea). Using 20 different ACPs for PCR, were cloned sequenced, and analyzed identities of 2 differentially expressed genes (DEGs). According to BLAST analysis, sequences of 2 clones significantly matched database entries and confirmed by semi-quantitative RTPCR. The functional roles of one up-regulated gene, cytochrome P450 mono-oxygenases 2K1v2 (CYP2K1v2), and one down-regulated gene was Profilin-1 were identified. We identified distinctive gene expression profiles in improved rainbow trout growth by treatment with a fish myostatin prodomain using ACP-based GeneFishing.

한국 주변해역 가리비로부터 분리한 18S rDNA 의 염기서열 분석

김미정(Mi Jung Kim),(Long Guo Jin),진형주(Hyung Joo Jin),조지영(Ji Young Cho),박중연(Jung Youn Park),장영진(Young Jin Chang),홍용기(Yong Ki Hong) 한국수산과학회 2001 한국수산과학회지 Vol.34 No.2

N/A Sequences of partial 18S rDNA have been analyzed to elucidate genetic diversity of scallops collected around Korea sea. The scallops used in genetic comparison are Argopecten irradians concentricus, Amusium japonicum japonicum, Chlamys farreri farreri, Chlamys(Swiftopecten) swifti and Patinopecten yessoensis. The 18S rDNA sequences were aligned by clustalx program. Phylogenetic tree was drawn by Treecon program. The scallops were divided into two groups-the Family Pectinidae containing A. japonicum japonicum and the Family Propeamussiidae containing Argopecten, Chlamys and patinopecten genera. The Family Propeamussiidae was also divided into the Supergenera Aequipecten containing A. irradians concentricus and Supergenera Chlamys containing C. farreri farreri, C. swifti and P. yessoensis. The species of C. swfti was closer to the P. yessoensis rather than C. farreri farreri in respect to nuclear 18S rDNA sequence.

제브라피쉬 근육성장에서의 carnosic acid의 효과

김정환 ( Jeong Hwan Kim ),진덕희 ( Diuk Hee Jin ),김영대 ( Young Dae Kim ),진형주 ( Hyung Joo Jin ) 한국어류학회 2014 韓國魚類學會誌 Vol.26 No.3

로즈마리의 주요 성분인 carnosic acid는 carnosol, rosmarinicacid, ursolic acid 등과 같은 폴리페놀의 한 성분으로 다양한 생리활성 기능이 보고되어 있다. 본 연구에서는 로즈마리 유래 폴리페놀인 carnosic acid가 제브라피쉬 근육성장에 미치는 영향을 근육 내 주사와 사료를 통해서 확인해 보았다. 근육 내 주사 실험을 통해서 CA는 제브라 피쉬의 근육 내 단백질 함량을 증가시키고 중성지방의 함량을 감소시켰다. 또한 조직학적 분석 결과 근섬유의 평균 면적이 커지는 근섬유의 과비대 효과를 나타내었다. 사료 실험 결과근육 내 단백질 및 중성지방의 함량에는 영향을 미치지 않았으며 조직학적 분석 결과 근육 내 주사 실험에서와 마찬가지로 근 섬유의 과비대를 유도하였다. Myogenesis is the formation process of multinucleated myofiber with a contractile capacity from muscle satellite cell (MSCs) during life. This process is tightly controlled by several transcription factors such as Pax3 and Pax7 (paired box protein 3 and 7), MEF2C (myocyte enhancer factor 2)and MRFs (myogenic regulatory factors) etc. On the contrary, myostatin (MSTN) is a transforming growth factor-β superfamily, which functions as a negative regulator of skeletal muscle development and growth. Carnosic acid (CA) is a major phenolic component in rosemary (Rosmarinus officinalis) and have been reported various biological activities such as anticancer, antioxidant, antimicrobial and therapeuticagents for amnesia, dementia, alzheimer`s disease. This study was confirmed to effects of CAon muscle cell line and muscle tissue alteration of zebrafish by intramuscular injection or feeding methods. 10 μM CA showed a non-cytotoxic on myoblast and a complete inhibition effect against myostatin activity on luciferase assay. In intramuscular injection experiment, the total protein and triglycerideamount of 10 μM/kg of CA injected group increased by 11% and decreased by 13% compared to these of the no injected group. In histology analysis of muscle tissues by hematoxylin/eosin staining, the number of muscle fiber of 10 μM/kg of CA injected group decreased by 29% and fiber area increased40% compared to these of no injected group. In feeding experiment, the total protein and triglycerideamount no significance difference compared to these of the normal feeding group. In histology analysis, the number of muscle fiber of 1% CA fed group decreased by 35% and fiber area increased 56%compared to these of normal fed group. We identified that CA have an effect on hypertrophy of musclefiber in adult zebrafish and the results of this study are considered as the basic data that can reveal the mechanisms of muscle formation via gene and protein level analysis.

어류 유래 마이오스타틴 프로도메인 단백질에 의한 시마연어(Oncorhychus masou) 성장효과

Jeong Hwan Kim(김정환),Sang Beum Lee(이상범),Mi Jin Cho(조미진),Ji Young Ahn(안지영),Suk Keun Lee(이석근),Sung-Youl Hong(홍성열),Ki Baik Seong(성기백),Hyung-Joo Jin(진형주) 한국생명과학회 2011 생명과학회지 Vol.21 No.8

성장과 분화를 조절하는 인자인 myostatin은 포유류에서 주로 골격근에 분포하며 근육성장을 억제하는 것으로 알려져 있다. Myostatin은 포유류에서뿐만 아니라 어류에 있어서도 그 기능이 유사하며 본 연구에서는 넙치와 조피볼락 유래 재조합 myostatin 단백질을 생산하여 시마연어에 침지방법을 통해 처리하였다. 처리 결과 시마연어의 무게와 생화학 분석에서는 유의성이 나타날 정도의 증가는 없었지만 근(muscle) 조직학적 분석에서 넙치와 조피볼락 유래 재조합 myostatin prodomain에 의해 12주째에는 세포의 수가 증가하는 hyperplasia가 일어났으며 22주째에는 조피볼락 유래의 재조합 myostatin prodomain을 처리한 군에서만 hypertrophy가 일어났다. 결론적으로 어류 유래 재조합 myostatin prodomain이 시마연어 근육성장 시 hyperplasia와 hypertrophy가 순차적으로 유도되는 것으로 확인되었다. Myostatin (MSTN) belongs to the transforming growth factor-β superfamily or growth and differentiation factor 8 (GDF-8), and functions as a negative regulator of skeletal muscle development and growth. Previous studies in mammals have suggested that myostatin knock-out increased muscle mass and decreased fat content compared to those of the wide type. Recently, several studies on myostatin have beenconducted on the block myostatin signal pathway with myostatin antagonists and the MSTN regulation with RNAi to control myostatin function. This study was performed to analyze growth and muscle alteration of Oncorhychus masou by treatment with recombinant myostatin prodomains derived from fish. We designed myostatin prodomains derived from P. olivaceus (pMALc2x-poMSTNpro) and S. schlegeli (pMALc2x-sMSTNpro) in a pMALc2x expression vector, and then purified the recombinant proteins using affinity chromatography. The purified recombinant proteins were treated in O. masou through an immersion method. Recombinant protein treated groups did not show a significant difference in weight, protein, or lipid composition compared to the control. However, there was a difference in the average number and area for histological analyses in the muscle fiber. At twelve and twenty-two weeks from the initial treatment, there were differences in average fiber number and area between the 0.05 ㎎/ℓ treated-group and the control, but the numbers were similar to those of the control during the same time period. At twelve weeks, however, 0.2 ㎎/ℓ treated-group had an increase in average fiber number and decrease in average fiber area compared to the control. At twenty-two weeks, the pMALc2x-sMSTNpro 0.2 ㎎/ℓ treated-group was induced and showed a decrease in average fiber number and increase in average fiber area. The results between twelve and twenty-two weeks showed that the fiber numbers had decreased, whereas average fiber area had increased due to sMSTNpro. It is understood that the sMSTNpro induced only hyperplasia at twelve weeks, after which it induced hypertrophy. Recombinant myostatin prodomains derived from fish may induce hyperplasia and hypertrophy in O. masou depending upon the time that has elapsed.

한국산 방사무늬김 (porphyra yezoensis)의 핵 18S rDNA 염기서열 분석

김명숙 ( Myung Sook Kim ),최재석 ( Jae Suk Choi ),조지영 ( Ji Young Cho ),진형주 ( Hyung Joo Jin ),홍용기 ( Yong Ki Hong ),( Long Guo Jin ) 한국수산과학회 2002 한국수산과학회지 Vol.35 No.6

N/A Nuclear 18S ribosomal RNA gene (18S rDNA) from the aquaculturable seaweed Porphyra yezoensis (Bangiales, Rhodophyta) was amplified using the polymerase chain reaction and its sequence was analyzed. Complete 18S rDNA has an 1823 bp exon and a 514 bp intron. The G+C contents of exon and intron were 48.6% and 55.4%, respectively. The exon sequence showed 99.5% homology to the GenBank accession number AB013177 of the Japanese P. yezoensis. The intron region that was inserted upstream between 568 and 1083 showed 93.4% homology to the AB013177.