• 생명과학회지
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• 제29권7호
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• pp.774-778
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• 2019

성숙 콩[Glycine max (L.) Merr.] 종자에는 단백질, 지방, 탄수화물의 3대 영양소와 아이소플라본 등 다양한 기능성 성분이 함유되어 있다. 그러나, Kunitz Trypsin Inhibitor (KTI) 단백질, 렉틴 단백질, 난소화성 올리고당인 stachyose 성분이 함유되어져 있어 품질과 기능성을 저하시키고 있다. 본 연구는 콩 및 콩 제품의 품질과 기능성을 저하시키는 KTI 및 렉틴 단백질과 stachyose의 성분 함량과 관련된 유전자들이 모두 열성으로 작용하는 콩 계통(triple recessive genotype)을 선발하기 위하여 진행되었다. 3개의 모본(개척2호, PI200508, 14G20)을 이용한 육종집단으로부터 성숙 종자에서KTI 및 렉틴 단백질이 없으면서 stachyose의 함량이 일반콩보다 현저히 적은 triple recessive 유전자형(titilelers2rs2) 개체를 선발하였다. 선발된 계통의 초장은 51 cm 정도였으며 백립중은 31.0 g으로 대립이었으며 종피색 및 제색은 노란색이었다. Stachyose의 함량(g/kg)은 일반콩(13.5 g/kg)보다 훨씬 낮았다(3.8 g/kg). 본 연구를 통하여 선발된 계통은 KTI 단백질과 렉틴 단백질이 동시에 없으며 stachyose의 함량이 낮은 다양한 콩 품종 육성을 위한 중간모본으로 이용될 수 있을 것으로 사료되었다.

• EDISON SW 활용 경진대회 논문집
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• 제5회(2016년)
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• pp.86-89
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• 2016

The Zika virus, which is a member of the flavivirus genus, poses a serious threat to humanity because there is no vaccine or cure. Zika is suspected to cause microcephaly, and it is rapidly spreading throughout parts of Brazil. Surprisingly, there are no known protein structures for the virus which are essential for drug and vaccine development. This paper investigates the Zika virus's nonstructural proteins with template-based modeling by using GalaxyTBM/Refine/SC. GalaxyDock was used to examine the effectiveness of various known serine protease inhibitors in inhibiting the Zika viral protease. In testing five inhibitors, Kunitz soybean trypsin inhibitor showed the strongest binding affinity (-10.082 kcal/mol). This paper provides a rudimentary foundation for further drug discovery research regarding the Zika virus.

• 한국작물학회지
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• 제57권1호
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• pp.78-82
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• 2012

인체내에서 소화불량 및 알레르기 반응을 일으킴으로써 콩의 품질을 저하시키는 물질인 Kunitz trypsin inhibitor(KTI) 단백질이 결핍되고, P34 단백질을 적게 함유하는 콩 계통을 선발하기 위해서, 현재까지 보고되지 않은 KTI 단백질의 유무와 P34 단백질 함량간의 유전관계에 대한 정보를 얻기 위하여 본 연구에서 얻어진 결과는 다음과 같다. 1. 07B1과 PI567476의 교배를 통해 얻어진 479개의 $F_2$ 종자를 대상으로 SDS-PAGE를 이용한 KTI 단백질의 유무를 확인한 결과, KTI 단백질이 존재하는 종자의 수는 353개, 결핍된 종자는 126개로 3 : 1로 분리하였다. 2 Western blot을 이용한 P34 단백질의 함량을 확인 한 결과, P34 단백질 함량이 보통 또는 높은 종자가 363개, 함량이 낮은 종자는 116개로 P34 단백질 함량에 대한 유전분리비는 3 : 1로 나타났다. 3. 전체 $F_2$ 종자 479개 중에서 KTI 단백질이 존재하며, P34 단백질 함량이 보통 또는 높은 종자가 266개, KTI 단백질이 존재하고 P34 단백질 함량이 낮은 종자가 88개, KTI 단백질이 결핍이고 P34 단백질함량이 보통 또는 높은 종자가 102개, KTI 단백질이 결핍이며 P34 단백질 함량이 적은 종자가 23개로 9:3:3:1의 분리비에 적합하여 KTI 단백질 유무와 P34 단백질 함량간에는 독립유전을 하였다.

• 한국육종학회지
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• 제42권1호
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• pp.35-39
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• 2010

Soybean proteins are widely used for human and animal feed in the world. ${\beta}$-conglycinin protein exhibits poor nutritional and food processing properties and Kunitz trypsin inhibitor (KTI) protein is a main anti-nutritional factor in soybean seed. The objective of this research was to identify the inheritance of $cgy_1$ gene and ti gene for the improvement of soybean cultivar with no KTI proteins and low amount of ${\beta}$-conglycinin. $F_2$ population was made by crossing between "Gaechuck2ho" (${\alpha}^{\prime}$-subunit present $Cgy_1Cgy_1$, KTI protein absent titi) and PI506876 (${\alpha}^{\prime}$-subunit absent $cgy_1cgy_1$, KTI protein present TiTi) parent. A total of 434 $F_2$ seeds were obtained and analyzed for the segregation of ${\alpha}^{\prime}$-subunit protein and KTI protein using SDS-PAGE. The segregation ratio of 3 : 1 for $Cgy_1$ locus (310 $Cgy_1$_ : 124 $cgy_1cgy_1$) and Ti locus (339 Ti_ : 95 titi) were observed. Segregation ratios of 9 : 3 : 3 : 1 (241 $Cgy_1$_Ti_: 69 $Cgy_1$_titi: 98 $cgy_1cgy_1$Ti_: 26 $cgy_1cgy_1titi$) between $Cgy_1$ gene and Ti gene in $F_2$ seeds were also observed (${\chi}^2= 5.367$, P = 0.10 - 0.20). This data showed that $Cgy_1$ gene was inherited independently with the Ti gene in soybean. These results will be useful in breeding program for selecting the line that does not exhibit or lacks both ${\alpha}^{\prime}$-subunit protein and KTI protein in soybean.

• Journal of Plant Biotechnology
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• 제45권4호
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• pp.328-332
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• 2018

Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrate, isoflavone and other various nutrients for humans and animals. However, there are anti-nutritional factors in the raw mature soybeans. Kunitz trypsin inhibitor (KTI) protein and stachyose are the main anti-nutritional factors in soybean seed. The ${\alpha}^{\prime}$-subunit of ${\beta}$-conglycinin protein exhibit poor nutritional and food processing properties. The genetic removal of the KTI and ${\alpha}^{\prime}$-subunit proteins will improve the nutritional value of the soybean seed. The objective of this research was to develop a new soybean strain with KTI and ${\alpha}^{\prime}$-subunit protein free ($titicgy_1cgy_1$ genotype) and proper agronomic traits. A breeding population was developed from the cross of the Bl-1 and 15G1 parents. A total of 168 $F_2$ seeds from the cross of the BL-1 and 15G1 parents were obtained. The segregation ratios of 9: 3: 3: 1 ($104Ti\_Cgy_{1\_}:\;30Ti\_cgy_1cgy_1:\;21cgy_1cgy_1Ti\_:\;13titicgy_1cgy_1$) between the Ti and $Cgy_1$ genes in the $F_2$ seeds were observed (${\chi}^2=5.12$, P=0.5-0.10). Two $F_4$ plant strains with proper agronomical traits and $titicgy_1cgy_1$ genotype (free of both KTI and ${\alpha}^{\prime}$-subunit protein) were selected and harvested. 2 strains (S1 and S2) had yellow seed coats and hilum. The plant height of the S1 strain was 65 centimeters. The 100-seed weight was 29.2 g. The plant height of the S2 strain was 66 centimeters and 100-seed weight was 26.2 g. The two strains selected in this research will be used to improve the new cultivar that will be free of the KTI and ${\alpha}^{\prime}$-subunit proteins.

• 생명과학회지
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• 제29권2호
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• pp.147-151
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• 2019

콩[Glycine max (L.) Merr. 2n=40]은 국내에서 벼 다음으로 많이 재배되고 있는 식량작물이며 성숙 콩 종실에는 3대 영양소외 페놀성 화합물, 이소플라본, 사포닌, 루테인 토코페롤, 피틴산, 루테인, 안토시아닌등 생리활성 물질이 다양하게 존재하지만 기능성과 가공 적성 및 품질을 저해시키는 성분들도 다수 존재한다. 성숙 콩 종실에서 품질, 기능성 및 가공 적성을 저하시키는 KTI 단백질이 없으면서 난소화성 올리고당인 stachyose 함량이 낮은 속 푸른 검정콩 유전자형을 육성하기 위하여 진행되었다. 3개의 모본으로 육종집단을 창성하여 검정종피, 녹색자엽 및 KTI 단백질이 없는 11개의 $F_2$ 종자를 얻었고 stachyose 함량이 낮은 2개의 계통을 선발하여 농업형질 평가를 통하여 검정 종피, 녹색자엽, stachyose 함량이 낮은 titirs2rs2 유전자형을 가진 계통을 선발하였다. 선발된 계통의 초장은 66 cm 정도였으며 백립중은 28.4 g으로 대립이었으며 종피색과 제색은 모두 검정색이었고 성숙 자엽색은 녹색이었다. 선발된 계통의 stachyose 함량은 일반품종(13.01-16.81 g/kg)보다 매우 낮은 2.59(g/kg)이었다. 본 연구를 통하여 선발된 계통은 Kunitz Trypsine Inhibitor (KTI) 단백질이 없으며 난소화성 올리고당인 stachyose 성분의 함량이 낮고 검정종피와 녹색자엽을 가진 기능성 고품질 유색콩 육성을 위한 중간모본으로 이용될 수 있을 것으로 사료되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제12권8호
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• pp.1251-1257
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• 1999

Two experiments were conducted to determine the effects of roasting and extrusion on nutritional value of conventional and low-inhibitor soy beans for nurser-age pigs. In Exp. 1, 100 weaning pigs (7.5 kg average initial BW) were used in a 35-d growth assay to determine the effects of processing method (roasting in a Rast-A-Tron$^{TM}$ raster vs extrusion in an Insta-Pro$^{TM}$ extruder) on the nutritional value of Williams 82 soybeans with (+K) and without (-K) gene expression for the Kunitz trypsin inhibitor. Treatments were 48% soybean meal with added soybean oil, +K roasted, +K extruded, -K roasted and -K extruded. All diets were formulated to contain 3.5 Mcal DE/kg, with 0.92% lysine for d 0 to 14 and 0.76% lysine for d 14 to 35 of the experiment. The lysine concentrations were 80% of NRC (1988) recommendations to accentuate difference in response to protein quality and lysine availability. For d 0 to 14, pigs fed extruded soybeans (+K and -K) had greater ADG (p<0.001), ADFI (p<0.09) and gain/feed (p<0.01) than pigs fed roasted soybeans. For d 14 to 35 and overall, the same effects were noted, i.e., pigs fed extruded soybeans had greater ADG, ADFI and gain/feed than pigs fed roasted soybeans (p<0.03). Also, pigs fed -K soybeans were more efficient (p<0.008) than pigs fed +K soybeans. In Exp. 2, 150 weanling pigs (7.0 kg average initial BW) were used in a 35-d growth assay. All diets were formulated to contain 3.5 Mcal DE/kg, with 1.25% lysine for d 0 to 14 and 1.10% lysine for d 14 to 35 of the experiment. The lysine concentrations were formulated to be in excess of NRC recommendation to determine if differences in nutritional value of the soybean preparations could be detected in protein-adequate diets. For d 0 to 14 (p<0.06), 14 to 35 (p<0.03) and 0 to 35 (p<0.02), pigs fed extruded soybeans had greater ADG and gain/feed than pigs fed roasted soybeans. Apparent digestibilities of DM, N and GE were greater for diets with extruded soybeans than diets with roasted soybeans and diets with soybean meal and soybean oil were intermediate. The response to extrusion processing was greater with -K than +K soybeans, with pigs fed extruded -K soybeans having the greatest growth performance and nutrient digestibilities and lowest skin-fold thickness of any treatment. In conclusion, extrusion yielded a full-fat soy product of greater nutritional value than roasting. Also, selection against genetic expression of the Kunitz trypsin inhibitor improved nutritional value of the resulting soybean preparations.

• BMB Reports
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• 제40권4호
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• pp.604-609
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• 2007

An Asp/His catalytic site of 10-formyltetrahydrofolate dehydrogenase (FDH) was suggested to have a similar catalytic topology with the Asp/His catalytic site of serine proteases. Many studies supported the hypothesis that serine protease inhibitors can bind and modulate the activity of serine proteases by binding to the catalytic site of serine proteases. To explore the possibility that soybean trypsin inhibitor (SBTI) can recognize catalytic sites of FDH and can make a stable complex, we carried out an SBTI-affinity column by using rat liver homogenate. Surprisingly, the Rat FDH molecule with two typical liver proteins, carbamoyl-phosphate synthetase 1 (CPS1) and betaine homocysteine S-methyltransferase (BHMT) were co-purified to homogeneity on SBTI-coupled Sepharose and Sephacryl S-200 followed by Superdex 200 FPLC columns. These three liver-specific proteins make a protein complex with 300 kDa molecular mass on the gel-filtration column chromatography in vitro. Immuno-precipitation experiments by using anti-FDH and anti-SBTI antibodies also supported the fact that FDH binds to SBTI in vitro and in vivo. These results demonstrate that the catalytic site of rat FDH has a similar structure with those of serine proteases. Also, the SBTI-affinity column will be useful for the purification of rat liver proteins such as FDH, CPS1 and BHMT.

• 한국작물학회지
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• 제54권3호
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• pp.299-306
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• 2009

콩과식물의 뿌리혹 형성을 조절하는 신호물질의 확인을 위해 신팔달콩2호의 줄기 수액 단백질 중에서 B. japonicum USDA110의 접종 후 2.5일(DAI)에 20 kDa의 SKTI 단백질이 증가하였다가 7 DAI에는 감소되면서 6 kDa의 작은 크기의 단백질이 증가되었다. 이러한 단백질의 차등발현은 조사한 3종의 콩에서 모두 유사하게 나타났으며 특히 대원콩에서 가장 두드러졌다. Western 분석으로 7 DAI에서 증가하는 6 kDa 단백질이 SKTI 항체와 특이적 반응을 하는 것으로 확인하여 SKTI가 절단되어 생긴 펩타이드로 추정되었다. 이러한 결과를 통해 20 KDa의 SKTI단백질이 콩의 뿌리혹 착생 초기단계인 2.5 DAI에 영향을 주고, 7 DAI로 진행되면서 6 kDa의 작은 크기의 단백질로 분해되어 그 양이 감소하는 것으로 생각된다. RNAi를 이용하여 유전자 기능이 억제된 형질전환된 모상근의 뿌리혹을 실제 형질전환이 확인된 모상근에 착생된 뿌리혹의 수를 비교한 결과 비재조합 A. rhizogenes을 접종시킨 대조구에 비해 SKTI RNAi 유전자를 형질전환한 모상근에서 모상근 당 착생된 뿌리혹 수가 감소되었다. 실시간 PCR 방법으로 형질전환된 모상근의 SKTI 전사체 수준에서도 상응하는 차이를 확인하였다. 이에 정확한 기작을 알 수 없지만 SKTI유전자가 뿌리혹 형성 초기에 뿌리혹 형성과정에 직간접적으로 관련하고 있음을 확인하였다. Sesbania rostrata의 뿌리혹 발생과정의 Protease 저해제와 같이 뿌리 혹 내의 감염세포 대 비감염세포의 비율을 조절하는 SKTI 발현 억제는 이러한 균형을 교란하여 뿌리혹의 생성을 억제하는 것으로 추정된다.

• 한국육종학회지
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• 제40권2호
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• pp.97-100
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• 2008

Lectin protein and Kunitz trypsin inhibitor (KTI) protein of mature soybean seed are a main antinutritional factor in soybean seed. The Le gene controls a lectin protein and Ti gene controls the KTI protein in soybean. Ti locus has been located on linkage group 9 in the classical linkage map of soybean. Position of Le locus on linkage map was not identified. Genetic relationship between Ti locus and Le locus could be useful in soybean breeding program for the genetic elimination of these factors. The objective of this study was to determine the independent inheritance or linkage between Ti locus and Le locus in soybean seed. Two $F_2$ populations were developed from three parents (Gaechuck#1, T102, and PI548415). The $F_1$ seeds from Gaechuck#1 (titiLeLe) ${\times}$ T102 (TiTilele) and Gaechuck#1 (titiLeLe) ${\times}$ PI548415 (TiTilele) were obtained. The lectin and KTI protein were analysed from $F_2$ seeds harvested from the $F_1$ plants to find independent assortment or linkage between Ti locus and Le locus. The segregation ratios of 3 : 1 for Le locus (129 Le_ : 44 lele) and Ti locus (132 Ti_ : 41 titi) and were observed. The segregation ratios of 9 : 3 : 3 : 1 (95 Le_Li_ : 34 Le_titi: 37 leleTi_ : 7 leletiti) between Le gene and Ti gene in $F_2$ seeds were observed. This data showed that Ti gene was inherited independently with the Le gene in soybean. These results will be helpful in breeding program for selecting the line with lacking both KTI and lectin protein in soybean.