• Journal of Microbiology and Biotechnology
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• 제17권5호
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• pp.728-732
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• 2007

In the present article, a novel fusion expression vector for Escherichia coli was developed based on the pTORG plasmid, a derivative of pET32a. This vector, named pT7MT(GenBank Accession No DQ504436), carries a T7 promoter and it drives the downstream gene encoding Metallothionein 2A(MT2A). There are in-framed multiple cloning sites(MCS) downstream of the MT2A gene. A target gene can be cloned into the MCS and fused to the C-terminal of the MT2A gene in a compatible open reading frame(ORF) to achieve fusion expression. The metal-binding capability of MT2A allows the purification of fusion proteins by metal chelating affinity chromatography, known as $Ni^{2+}$-affinity chromatography. Using this expression vector, we successfully got the stable and high-yield expression of MT2A-GST and MT2A-Troponin I fusion proteins. These two proteins were easily purified from the supernatant of cell lysates by one-step $Ni^{2+}$-affinity chromatography. The final yields of MT2A-GST and MT2A-Troponin I were 30mg/l and 28mg/l in LB culture, respectively. Taken together, our data suggest that pT7MT can be applied as a useful expression vector for stable and high-yield production of fusion proteins.

• 미생물학회지
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• 제41권3호
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• pp.195-200
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• 2005

Comamonas sp. strain DJ-12의 pmcABCDEFT 유전자군은 protocatechuate (PCA)의 분해과정에 관여하는 PCA 4,5-dioxygenase, 4-carboxy-2hydroxymuconic semialdehyde (CHMS) dehydrogenase, 2-pyrone04,5-dicarboxylate(PDC) hydrolase, 4-oxalomesaconate (OMA) hydratase, 그리고 4-oxalocitramalate (OCM) aldolase 등의 효소들을 생산하는 유전자들과 transporter의 역학을 하는 유전자로 각각 확인되었다. 이 유전자군은 Comamonas sp. strain DJ-12의 chromosomal DNA로부터 얻은 PCR 산물들을 T-vector에 ligation하여 재조합 플라스미드 pMT1, pMT2, pMT3, pMT4, pMT5, pMT6, pMT7, pMT8, pMT9, pMT10을 제조하였다. 이들 재조합 플라스미드의 염기서열을 분석한 결과 PCA 4,5-dioxygenase 유전자는 alpha(pmcA)와 beta(pmcB) 두 개의 subunit으로 구성 되어있으며, 각각 450 bp와 870 bp이었다. CHMS dehydrogenase 유전자(pmcC)는 960 bp, PDC hydrolase 유전자(pmcD)는 918 bp이였으며, OMA hydratase 유전자(pmcE)는 1029 bp, OCM aldolase 유전자 (pmcF)는 689 bp, 그리고 transporter 유전자(pmcT)는 1,398 bp이였다. 이들 pmc 유전자들은 pmcT-pmcE-pmcF-pmcD-pmcA-pmcB-pmcC의 순서로 배열되어 있었다. Comamonas sp. strain DJ-12의 pmcABCDEFT 유전자산물의 아미노산 서열을 분석한 결과, Comamonas testosteroni BR6020 및 Psedomonas ochraceae NG.J1와 $94{\~}98\%$의 높은 유사성을 보였고, 그 유전자들의 배열 순서도 동일하였다. 그러나 Sphingomonas paucimobilis SYK-6, Sphingomonas sp. LB126, 그리고 Arthrobacter keyser 12B와는 아미노산 서열이 $52{\~}74\%$의 유사성을 보였고, 그 유전자의 배열 구조도 상이하였다.

• 한국축산식품학회지
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• 제25권2호
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• pp.218-225
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• 2005

본 연구는 mt DNA의 D-loop 영역을 probe로 이용한 Southern blot hybridization 분석 기법을 이용하여 한우에서 mt DNA의 RFLP를 분석하고 RFLP marker가 유생산에 미치는 영향을 분석하여 산유량 관련 DNA marker를 개발하고자 수행하였다. Mt DNA의 D-loop 영역내 404번부터 15061까지 1142 bp 크기의 염기 서열 부위를 특이적인 primer를 이용하여 PCR로 증폭하였다. Mt DNA를 HpaII, BamHI, XbaI, HinfI, EcoRI, HindII 및 RsaI 7종류의 제한효소를 이용하여 각각 절단한 후 DIG로 표지된 D-loop probe를 이용하여 검출한 결과 XbaI, RsaI, BamHI 및 HpaII 4종류의 제한효소에서 각각 RFLP 다형성이 검출되었고 EcoRI, HindIII 및 HinfI 3종류 제한효소는 변이가 존재하지 않았다. 다유 계통과 저유 계통 선발 집단간의 각 제한효소별 RFLP type의 출현빈도를 비교한 결과 BamHI 및 RsaI 제한효소에서 두 집단간의 RFLP type의 출현율에 각각 통계적 유의성(P<.05)이 인정되었다. 다유 및 저유성으로 극단의 육종가 값을 갖는 두 계통의 mt DNA D-loop영역의 염기 서열을 비교 분석한 결과 441번째 염기가 G/C, 469번째 염기는 T/C, 503번째 염기는 C/T, 569번째 염기는 G/A, 614번째 염기는 C/A그리고 644번째 염기는 C/T로 각각 염기가 치환되었고 특히, 다유 계통 개체의 677번째의 A염기가 저유 계통 개체에서는 결실되어 있다는 사실이 확인되었다. 한편, 한우 암소의 유생산에 영향을 미치는 효과를 규명하기 위하여 mt DNA RFLP 형과 송아지 이유시 체중, 생시체중 및 비유량을 측정하여 얻어진 육종가의 성적을 근거로 통계 분석한 결과 XbaI 제한효소의 RFLP type이 산유 능력 육종가와 유의적인 관련성이 확인되었다 (P<05). 즉, RFLP A type을 갖는 축군의 평균 육종가 추정치가 6.233으로 B type을 갖는 축군의 평균 육종가 추정치 0.757보다 월등히 높은 것으로 나타났다. 결론적으로 산유량과 관련성이 확인된 mt DNA RFLP type은 한우의 산유량 향상을 위한 DNA marker로 이용 가능할 것으로 기대된다.

• 한국유기농업학회지
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• 제8권3호
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• pp.99-109
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• 2000

제주지방에서 가을감자의 더뎅이병 방제 및 수량증대를 위해 전작물로 녹비작물(콩, clover, pioneer, orchardgrass)을 '98년 5월 21일부터 7월 28일까지 재배하여 생육최성기에 파쇄, 경운하고 P.E 투명필름(0.01mm)으로 멀칭후 터널 설치, 태양열 토양소독을 실시한 포장에 가을감자를 재배하여 생육상태를 조사한 결과는 다음과 같다. 1. 전작물로 재배한 청예수량은 pioneer BSSF(64.3MT/ha)가 가장 많았으며, 다음으로 콩((25.0MT/ha), clover, orchardgrass 순이었다. P.E 필름멀칭+터널설치중의 상승한 평균지온은 토양깊이 5cm에서 54$^{\circ}C$, 10cm에서 45$^{\circ}C$, 20cm에서 44$^{\circ}C$였다. 콩, red clover 재배구가 비닐멀칭에 의한 토양소독효과로 인하여 가을감자의 더뎅이병 발병도가 낮아지는 것으로 조사되었다. 2. 초장, 엽록소함량치, 지상부생체중, 괴경수량 등의 생육형질은 녹비작물재배 및 비닐멀칭처리에 의해 양호해지는 경향을 보였다. 콩, red clover, pioneer 855F 재배후 감자재배구가 괴경수량이 많아지는 것으로 조사되었다. 3. 가을감자 지상부 및 괴경 중의 T-N, K, Ca함량도 녹비작물재배 및 P.E 필름멀칭에 의해 높아지는 경향을 보였다. 반면에 토양중의 T-N, 유기물함량, K함량은 녹비작물재배 및 P.E 필름멀칭후에 높아졌다가 가을감자 수확후에는 시험시작전 토양의 상태로 다시 낮아지는 추세를 나타내었다

• Korean Journal of Radiology
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• 제21권5호
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• pp.588-597
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• 2020

Objective: To investigate the value of combined chemical exchange saturation transfer (CEST) and conventional magnetization transfer imaging (MT) in detecting metabolic and structural changes of renal fibrosis in rats with unilateral ureteral obstruction (UUO) at 3T MRI. Materials and Methods: Thirty-five Sprague-Dawley rats underwent UUO surgery (n = 25) or sham surgery (n = 10). The obstructed and contralateral kidneys were evaluated on days 1, 3, 5, and 7 after surgery. After CEST and MT examinations, ¹⁸F-labeled fluoro-2-deoxyglucose positron emission tomography was performed to quantify glucose metabolism. Fibrosis was measured by histology and western blots. Correlations were compared between asymmetrical magnetization transfer ratio at 1.2 ppm (MTR_asym(1.2ppm)) derived from CEST and maximum standard uptake value (SUV_max) and between magnetization transfer ratio (MTR) derived from MT and alpha-smooth muscle actin (α-SMA). Results: On days 3 and 7, MTR_asym(1.2ppm) and MTR of UUO renal cortex and medulla were significantly different from those of contralateral kidneys (p < 0.05). On day 7, MTR_asym(1.2ppm) and MTR of UUO renal cortex and medulla were significantly different from those of sham-operated kidneys (p < 0.05). The MTR_asym(1.2ppm) of UUO renal medulla was fairly negatively correlated with SUV_max (r = -0.350, p = 0.021), whereas MTR of UUO renal medulla was strongly negatively correlated with α-SMA (r = -0.744, p < 0.001). Conclusion: CEST and MT could provide metabolic and structural information for comprehensive assessment of renal fibrosis in UUO rats in 3T MRI and may aid in clinical monitoring of renal fibrosis in patients with chronic kidney disease.

• Journal of Ecology and Environment
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• 제29권5호
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• pp.439-443
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• 2006

Soil properties of Quercus variabilis forest on Youngha valley at Mt. Worak National Park were studied as a part of Korea National Long-Term Ecological Research. Soil sampling was carried out along the 50 cm soil depth with 10cm intervals at every quarter from May 2005 through July 2006. Fresh soil was used for $NH_4{^+}-N,\;NO_3{^-}-N$, and soil water content determination. Remaining soils were air dried in the shade, and then used for determination of soil pH, T-N, T-P and exchangeable cation. Average soil organic matter in top soil was $8.5{\pm}1.2%$ and decreased with soil depth. Bulk density of top soil was $0.82{\pm}0.07g/cm^3 $and increased with soil depth. Soil organic matter and bulk density showed a negative linear correlation ($R^2=0.8464$). Soil pH in top soil and subsoil was similar. T-N, $NH_4{^+}-N,\;NO_3{^-}-N$ and T-P in top soil were $1.9{\pm}0.5mg/g,\;7.3{\pm}1.0mg/kg,\;2.0{\pm}0.4mg/kg\;and\;0.2{\pm}0.05mg/g$, respectively. $K^+,\;Ca^{2+}\;and\;Mg^{2+}$ in top soil were $84.6{\pm}24.4,\;408.8{\pm}137.8\;and\;93.4{\pm}23.0mg/kg$, respectively. They decreased with soil depth. Amounts of organic matter, T-N, $NH_4{^+}-N,\;NO_3{^-}-N$, T-P, $K^+,\;Ca^{2+}\;and\;Mg^{2+}$ in 50 cm soil depth were 250.9, 3.45, 0.025, 0.003, 0.639, 0.181, 0.845 and 0.302 ton $ha^{-1}\;50cm-depth^{-1}$, respectively.

• Mycobiology
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• 제50권5호
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• pp.317-325
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• 2022

Endophytic fungi occupy various ecological niches, which reinforces their diversity. As few studies have investigated the endophytic fungi of alpine conifers, we focused on four species of alpine conifers in this study-Abies nephrolepis, Pinus pumila, Taxus cuspidata var. nana, and Thuja koraiensis-and examined them for endophytic fungi. A total of 108 endophytic fungi were isolated. There were four taxa in A. nephrolepis, 12 in P. pumila, 18 in T. cuspidata var. nana, and 17 in T. koraiensis; these were divided into five classes: Agaricomycetes (3.2%), Dothideomycetes (29.0%), Leotiomycetes (15.0%), Sordariomycetes (41.9%), and Orbiliomycetes (1.6%). The most prevalent fungi were Sydowia polyspora (22.7%) and Xylariaceae sp. (22.7%) in P. pumila, Phomopsis juglandina (16.1%) in T. cuspidate var. nana, and Thuja-endophytes sp. 1 (70.0%) in T. koraiensis. However, there was no dominant species growing in A. nephrolepis. Some host plants were analyzed using next-generation sequencing. We obtained 4618 reads for A. nephrolepis and 2268 reads for T. koraiensis. At the genus level, the top three endophytic fungi were Ophiostomataceae_uc (64.6%), Nectriaceae_uc (15.5%), and unclassified organism (18.0%) in A. nephrolepis and Nectriaceae_uc (41.9%), Ophiostomataceae_uc (41.8%), and Magnaporthaceae_uc (9.2%) in T. koraiensis. Our results show that there are different communities of endophytic fungi among different host plants, even if the host plants are in the same region. Such ecological niches are important in terms of the ecological restoration of alpine conifers.

• 한국응용과학기술학회지
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• 제18권3호
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• pp.197-213
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• 2001

Conjugate trienoic acids (CTA) occurred in triacylglycerols (TGs) of the seed oils of Trichosanthes kirilowii, Momordica charantia and Aleurites fordii, and they were easily converted to their methyl esters in a mixture of sodium methoxide-methanol without any structural destruction. The main fatty acids in triacylglycerol (TG) fraction of the seed oils of Trichosanthes kirilowii are $C_{18:2{\omega}6}$ (32.2 mol %), $C_{18:3{\;}9c.11t,13c}$ (38.0 mol %) and $C_{18:1{\omega}9}$ (11.8 mol %), followed with $C_{16:0}$ (4.8 mol %) and $C_{18:0}$ (3.1 mol %). The TG fraction was resolved into 20 TG molecular species according to the partition number (PN) by reversed-phase (RP)-HPLC. The main TG species were $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$, of which amounts reached 63 mol % of total TG molecular species. The TG sample was fractionated into 11 fractions according to the number of double bond in the molecule by $Ag^{+}-HPLC$ and the species of $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$ were also eluted as main components. The TG species containing CTA showed unusual behaviours in the order of elution by HPLC ; first, TG moleular species of $DT_{c2}$ (D; dienoic acid, $T_{c}$; punicic acid, $T_{ci}$; ${\alpha}-eleostearic$ acid, M ; monoenoic acid, $S_{t}$; stearic acid) was eluted earlier than $Mt_{c2}$, although they have the same PN number of 40, and, secondly, the species of $DT_{ci2}$ with eight double bonds was eluted earlier than that of $D_2T_{ci}$ with seven double bonds. Intact TG of the seed oils of Momordica charantia contained mainly fatty acids such as $C_{18:3{\omega}9c,11t,13t}$ (57.7 mol %), $C_{18:1{\omega}9}$ (17.4 mol %), $C_{18:0}$ (12.3 mol %) and $C_{18:2{\omega}6}$ (10.6 mol %), and was classified into 13 fractions by RP-HPLC. The main TG species were as follows ; $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.1 mol %] and $S_{t}T_{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_2$, 33.9 mol %] comprising about 73 mol % of total TG species, accompanied by $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 7.3 mol %], $D_{2}T_{ci}$ [$ (C_{18:2{\omega}6})_{2}(C_{18:3\;9c,11t,13t})$, 3.6 mol %] and $MDT_{ci}$ [$(C_{18:1{\omega}9})(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})$, 3.5 mol %]. Simple TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_3]$ was present in a small amount of 1.4 mol %, but other simple TG species were not detected. The TG was also resolved into 11 fractions according to the number of double bond by $Ag^{+}-HPLC$, and the species were mainly occupied by $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.4 mol %] and $S_tT-{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_{2}$, 35.4 mol %] $DT_{ci2}$ species with eight double bonds was also developed faster than $D_2T_{ci}$ one with seven double bonds as indicated in the analysis of TG of the seed oils of T. kirilowii, and $MT_{ci2}$ species with cis, trans, trans-configurated double bond was eluted earlier than $MT_{c2}$ species with cis, trans, cis-configurated double bond. The main components of fatty acid in total TG fraction isolated from the seed oils of of Aleurites fordii were in the following order ; $C_{18:3\;9c,11t,13t}$ (81.2 mol %)> $C_{18:2{\omega}6}$ (8.5 mol %)> $C_{18:1{\omega}9}$ (5.4 mol %)$. With resolution of the TG by RP-HPLC, eight fractions such as $T_{ci3}$, $Dt_{ci2}$, $D_{2}T_{ci}$, $MT_{ci2}$, $PT_{ci2}$ (P; palmitic acid), $PMT_{ci}$, $PDT_{ci}$ and $S_{t}T_{ci2}$ ($S_{t}$; stearic acid) were isolated, respectively. TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_{3}$, 54.2 mol %], $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 15.0 mol %] and $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3 9c,11t,13t})_{2}$, 14.8 mol %] were present as main species.

• 한국습지학회지
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• 제23권3호
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• pp.252-259
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• 2021

무등산에서 발원하여 광주천으로 유입되던 수많은 계곡수가 합류식 하수관거로 유입되면서 광주천의 건천화와 수질 악화가 심화되었다. 본 연구는 하수처리장으로 유입되는 광주천 유역의 무등산 계곡수를 수원으로 활용하여 실개천을 조성하는 방안에 대해 연구하였다. 먼저, 100 m³/일 이상의 유량을 보이는 4개 지점의 유량 및 수질 조사 결과 지점별로 105~2,721 m³/일의 유량을 나타냈으며, 평균 수질은 BOD₅ 0.3~1.6 mg/L로 나타났다. 만약, 갈수기 유량 1,500 m³/일의 실개천을 조성한 후 광주천 남광교 지점으로 유입시킨다면, BOD 7.3%, COD 6.5%, T-P 5.8%, T-N 5.2%의 개선 효과가 있을 것으로 예측되었다. 또한, 계곡수 유입에 의한 하수처리장 유량 부하 부담이 줄어들고, 하수처리 비용이 절감되며, 도심 속 수변 amenity 창출에 의한 Blue-Green-Network (BGN) 구축의 토대가 될 것으로 판단되었다.

• 수산해양기술연구
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• 제44권4호
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• pp.282-293
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• 2008

In order to develop the substitutive materials for natural baits of swimming crab pots, the attracting effects of swimming crab such as the preference of baits which were made of the by-products of marine and stock raising through the water tank experiments and fishing experiments. On the investigation of mean entrapped catch number to the pot by the baits after putting the 4 kinds of baits, mackerel(M), mackerel with grinded mackerel s internals($MM_I$), mackerel with tuna s internals$MM_I$) and makerel with grinded krill(MK) each in one pot by turns, $MM_I$ and MK were entrapped mean 3.9(13.0%) and they were a little more comparing to M, and $MT_I$ is least with mean 2.1(7.0%)(F=12.913, P < 0.05). Otherwise, on the preference investigation of swimming crabs by the baits after putting the 4 kinds of baits in the 4 pots each, M was entrapped mean 3.0(10%), but $MM_I$, $MT_I$ and MK were mean 1.2(4.0%), 1.0(3.3%) and 1.5(5.0%) each and they were only 30-50% of M(F=13.398, P < 0.05). On the preference investigation of swimming crabs by the 5 kinds of baits, mackerel(M), and krill(K), manila clam($M_C$), pig s fat($P_F$) and chicken s head($C_H$) which were used in substitutive baits, M was entrapped mean 3.2(10.7%), but K was about 50% of catch of M with mean 1.6(5.3%), and $M_C$, $P_F$ and $C_H$ were very few with mean 0.1-0.2(0.3-0.7%)(F=89.186, P < 0.05). On the preference investigation of swimming crabs by the pots which were put each the 3 kinds of baits, original krill(K), grinded krill with gluten and soybean oil cake($K_GGS$) and grinded krill with gluten, soybean oil cake and glycine($K_GGSG_L$) in the blue fluorescent hexahedral plastic bait cages(BF), and which were put the mackerel(M) in the non-fluorescent hexahedral red plastic bait cage($RF_N$), it was entrapped mean 3.0(10.0%) in the pot which was put the mackerel in the $RF_N$, and the same level in the pots which were put the K and $K_GGSG_L$ in the BF, but it was mean 2.0(6.7%) in the pots which was put the $K_GGS$ in BF and it was decreased by 30% of catch comparing to $RF_N$(F=3.750, P < 0.05). On the preference investigation of swimming crab by the pots which was put grinded tuna with gluten, soybean oil cake and glycine($T_IGSG_L$) in the blue fluorescent hexahedral plastic bait cage(BF), and which was put mackerel(M) in the nonfluorescent hexahedral red plastic bait cage($RF_N$), it was entrapped mean 3.3(11.0%) in the pot which was put mackerel in $RF_N$, and mean 2.7(9.0%) in the pot which was put $T_IGSG_L$ in BF and it was about 15% less comparing to use bait M(t=1.387, P < 0.05). As a results of fishing experiments, a plan for enhancing catching efficiency of $T_IGSG_L$ will be required because catching efficiency of $T_IGSG_L$, alternative bait, was half of fish catching efficiency of natural bait using mackerel. Fishing experiments were conducted 3 times using reinforced substitutive artificial bait that is reinforced attractive effect of $T_IGSG_L$ and composed of tuna intestine, grinded mackerel, gluten, soybean cake, glycine and alanine($T_IM_GGSG_LA$). Catching efficiency of $T_IM_GGSG_LA$ was about 80% of that of natural bait made of mackerel.