핵다면체 바이러스 DNA 단편의 클로닝과 발현 및 바이러스 발현 운반체 제조

이근광(Keun-kwang Lee) 군산대학교 수산과학연구소 1993 水産科學硏究 Vol.- No.9

HcNPV DNA genome을 제한효소 EcoR Ⅰ으로 절단하고, pUC8 plasmid vector를 사용하여 E coli JM 83세포에 형질전환하였다. 4개의 EcoR Ⅰ DNA 단편을 pUC8 plasmid vector의 EcoR Ⅰ 부위에 cloning하고, 그들 DNA를 다시 EcoR Ⅰ으로 절단하여 확인하였다. 이들 재조합체는 각각 LK-1, LK-2, LK-3, LK-4로 명명하였다. 재조합체와 E coli JM83 세포의 전체 단백질 polypeptide를 SDS-AGE로 분석하였다. 그 결과 재조합체 LK-1, LK-2, LK-3는 E coli JM83의 단백질 밴드와는 달리 하나의 새로운 밴드가 나타났다. 새로 나타난 밴드의 분자량은 약 47.5kd이다. 또한, HcNPV DNA 게놈을 제한효소로 절단한 단편상에서 cDNA probe와 혼성화시킨 결과 Kpn Ⅰ은 A 단편(54.5kb), Pst Ⅰ은 F 단편(7.6kb), Pvu Ⅱ은 L 단편(93.34kb) 그리고 Xho Ⅰ은 B와 C 단편(28.4와 23.1kb)에서 양성 반응을 보였다. polyhedrin 유전자의 일부 promoter 부위의 염기서열을 결정하였다. 그 결과 TATA box는 전사 개시 위치로부터 위쪽으로 약 21bp의 polyhedrin 유저자 5' flanking 부위에서 발견되었다. polyhedrin 유전자내 CAAT는 TATA-like box측면 염기에 존재하지 않았다. 또한 4개의 앞뒤로 반복되는 5'-CTAATAT-3와 5'TAAATAA-3'의 염기는 polyhedrin 유전자내 전이 개시 위치로부터 위쪽으로 83과 50과 25bp부위에 존재하였으며, 다른 하나는 전이 개시 위치로부터 아래쪽으로 4bp부위에서 발견되었다. 그리고 polyhedrin 유전자내 전이 개시 위치로부터 위쪽으로 약 83bp부위는 AT(78%)가 매우 많이 존재하였다. Bculovirus 발현 운반체를 구성하기 위해서 pUC8 운반체를 이용하였으며, pUC8운반체의 EcoR Ⅰ 부위를 삽입하여 운반체를 구성하였고, 이를 pBE4라 명명하였다. 또한 pBE4 clone DNA의 cloning 부위중 BamH Ⅰ과 Hind Ⅲ 부위를 이중 절단한 후, 1.1kb의 bovine growth hormone 유전자를 삽입 연결 하였고, 이를 pbGHL1이라 명명하였다. 재조합 바이러스를 제조하기 위하여 pbGHL1 plasmid DNA와 HcNPV DNA를 S. frugiperda 세포에 동시 형질감염시켜 재조합 바이러스를 얻었고, 이를 pbHcL1이라 명명하였다. Bovine growth hormone을 방사성 면역측정 방법으로 측정하였다. 그 결과 pbGHL1은 0.09ng/ml이 생산되었으며, pbHcL1은 0.07ng/ml이 생산되었다. Baculovirus, H cunea nuclear polyhedrosis virus(NPV) DNA genome was digested with EcoR I endonuclease and transformation in E. coli JM 83 using the plasmid vector pUC8. Four EcoR I site of plasmid vector pUC8 and identified by digestion of the recombient DNAs with EcoR I enzyme. The recombinents are named as LK-l, LK-2, LK-3 and LK-4. The total cell polypeptides of the recombinents and E. coli JM 83 were analyzed on the SDS-PAGE. Recombinents, LK-l, LK-2, and LK - 3 were appeared one new band as comparison with polypeptide bands of E. coli JM 83 cell. The molecular weight of new band was approximately 47.5Kd. The DNA genome was digested with Kpn Ⅰ, Pst Ⅰ, Pvu Ⅱ and Xho Ⅰ restriction endonucleases and hybridized with (a^(32)p)-labelled AcNPV polyhedrin gene cDNA. The probe DNA hybredized to Kpn Ⅰ-A, Pst Ⅰ- F, Pvu Ⅱ-L and Xho Ⅰ-B, C segment. The promoter region of polyhedrin gene of HcNPV genome DNA was sequenced. The sequences identified to the TATA box was found at the 5' flanking region of the polyhedrin gene approximately 21 bp upstream from the transcriptional start site. But CAAT-like box in the polyhedrin gene was not found in the sequences flanking the TATA-like box. Two fragments of tandem repeats with the sequence 5'-CTAATAT-3' and 5'-TAAATAA-3' were found between 83 and 50 or 25 upstream and 4 bp downstream from the translation start site. About 83 bp region upstream from the translational start site was highly AT(78%) rich. pUC8 plasmid vector and EcoR Ⅰ-H fragment were digested with BamH Ⅰ/EcoR Ⅰ and ligated to be named as pBE4. Bovine growth hormone gene(1.1kb) was inserted into BamH Ⅰ and Hind Ⅲ site of the pBE4 and named as pbGHL1. For the construction of recombinant viral DNA, pbGHL1 plasmid DNA and HcNPV DNA were cotrasfected S. frugiperda cell and selected polyhedra negative plaque, which was named as pBHcL1. Bovine growth hormone production was measured by radioimmunioassay method. The amount of bovine growth hormone produced by pbGHL1 was 0.09ng/ml and 0.07ng/ml in pbHcL1 respectively.

기능성 화장품 소재로서 금잔화 꽃 추출물의 조성 및 항산화 활성 연구

이근광 ( Keun Kwang Lee ),권호선 ( Ho Sun Kwon ),송연숙 ( Yeon Sook Song ),김명곤 ( Myung Kon Kim ) 한국미용학회 2015 한국미용학회지 Vol.21 No.6

This study was to investigate the phenolic compound compositions and DPPH free radical scavenging activity, and possibility of beauty cosmesutical ingredients of Calendula arvensis flower extracts. The results were as follows: In term of the free amino acid of the C. arvensis flower extract obtained using hot water and 80% MeOH, a total of 17 different compositions were obtained. In the hot water extract, 5.16 mg/g was detected. Among them, arginine was the highest with 0.72 mg/g. However, total contents in 80% MeOH were 4.41 mg/g. Among them, serine(0.54 mg/g) was the highest. In the polyphenol contents, 80% MeOH extract was greater than hot water extract. In terms of flavonoid contents, 80% MeOH extract was about 5 times greater than hot water extract. Regarding the composition of phenolic compound, 7 different types of phenolic acids and zero types of flavonoids were found in hot-water. And 7 different types of phenolic acids and one types of flavonoids were found in methanol extracts. The DPPH free radical scavenging activity was 66.99 ± 0.67% and 77.63 ± 0.48% in the hot water extract and 80% MeOH extracts respectively. With high DPPH free radical scavenging activity, the possibility as a natural antioxidant was high. In the inhibitory activity of tyrosinase, 80% MeOH extracts was about 1.3 times greater than hot water extract. When the C. arvensis flower hot water extract was treated to the human keratinocyte cell line HaCaT, cells grew in proportion to the concentration. At 100 μg/ml, for example, it increased by about 28%, compared to the control group. In the 80% MeOH extract, on the other hand, almost no cytotoxicity was found until 100 μg/ml.

식물 추출 정유가 어류의 병인체에 미치는 항미생물 효과

이근광(Keun-Kwang Lee),이정열(Jeong-Yeol Lee) 한국수산과학회 양식분과 2001 韓國養殖學會誌 Vol.14 No.2

품바의 미용분장 특성 및 토탈 코디네이션 연구

이근광 ( Keun-kwang Lee ) 대한미용문화예술학회 2017 대한미용문화예술학회지 Vol.6 No.1

The following are characteristics of the face makeup and the total co-ordination according to the characteristics of the pumba and the road festival. The pumba troupe is generally paired with five to six couples, dressed in various funny dresses, costumes, accessories and performances by singing and dancing. As a result, they are invited to festivals in each region, group events at the end of the year, events in the house and not only create a mood with songs, dances and daily life humor but also help to relieve stress. They are also a type of profession that receives donations from spectators in exchange for performances, or sells traditional candy or household goods. Their face make-up characteristics in the streets are full of make-up on the face and eyebrows are painted neatly along the eyebrows. their face are painted with reddish paper on both sides of the face, menggujeom on her nose as a point, reddish paper on her nose as well to express strawberry, and red lipstick on her lips to express humorous appearance. In addition, the total coordination of the Pumba performance team, such as hair, costumes, accessories, and performances, features a variety of hair ornaments and flowers, and hair styles such as using a rubber band to bind. The accessories include earrings, rings, sales, hair clippers, etc. and the performance of various kinds of drums, hourglass, gong, yeojjangsu, etc. is used to enhance the fun. Therefore, when combined with these results, pumba can not only relieve stress by giving laughter to crowd as a result of performing practical acts, such as laughter, comedy, and costume coordinators, but also show that the performance of the musical is one of the regional cultural arts. Therefore, it is expected that the interest and support will be actively needed so that the performers will continue to understand the hardships and pain of the people who are more likely to understand the hardships and pain of the people and continue the performances that will give them joy.

A Infectious Diseases Control Multi-Agent System using Artificial Intelligence Learning Algorithm

Keun-Kwang Lee(이근광),Hee-Sook Kim(김희숙),Min-Hi Lee(이민희) 한국디지털콘텐츠학회 2019 한국디지털콘텐츠학회논문지 Vol.20 No.10

모바일 두피 진단 시스템에 관한 연구

이근광(Keun-kwang Lee),김희숙(Hee-Sook Kim),이민희(Min-Hi Lee) 한국디지털콘텐츠학회 2019 한국디지털콘텐츠학회논문지 Vol.20 No.11

양식 가물치 ( Channa argus ) 에 대한 Saprolegnia sp. 의 병리학적 특성과 물곰팡이의 생장을 제어하는 정유의 영향

이근광(Keun Kwang Lee),김영길(Young Gill Kim),이민웅(Min Woong Lee),이형환(Hyung Hoan Lee) 한국균학회 1999 韓國菌學會誌 Vol.27 No.1

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Electron Microscopic Visualization of Dendrolimus spectabilis Midgut Cells infected by D. spectabilis Nuclear Polyhedrosis Virus

이형환,이근광,이민웅,Lee, Hyung-Hoan,Lee, Keun-Kwang,Lee, Min-Woong Korean Society of Electron Microscopy 1991 Applied microscopy Vol.21 No.1

Dendrolimus spectabilis nuclear polyhedrosis virus(DsNPV)에 감염되어 죽은 솔나방(D. spectabilis)유충의 중장세포를 전자현미경(TEM)으로 관찰하였다. 감염된 중장세포의 핵에서 DsNPV는 복제하였으며, DsNPV의 virogentic stroma가 핵속에 나타났고, nucleocapsid도 형성되었다. 또한 NPV의 polyhedra형성을 핵에서 관찰했다. Midgut cells of the dead Dendrolimus spectabilis larvae by infection of D. spectabilis nuclear polyhedrosis virus (DsNPV) were observed by transmission electron microscopy. DsNPV replicated in the nucleus of the infected midgut cells and the virogenic stroma of DsNPV appeared in the nucleus, from which nucleocapsids were formed. Also the formation of polyhedral inclusion bodies were observed in the nucleus.

여성의 외적이미지 관심도와 헤어스타일에 따른 특성 연구

이옥순 ( Ok-soon Lee ),송연숙 ( Yeon-sook Song ),이근광 ( Keun-kwang Lee ) 대한미용문화예술학회 2018 대한미용문화예술학회지 Vol.7 No.2

This study analyzed interest of women’s appearance image and characteristics according to hair style. For the research method, this study used surveys and the subjects were 276 women from Daejeon and Chungnam. The collected data were analyzed using SPSS(Statistical Package for Social Science) 24.0 and the results are as follows. According to the survey, age over 50s of respondents was topped the list with 32.2%. The majority(69.9%) were married and also 27.7% said their monthly income was less than 4 million won. 64.5% were undergraduate or graduate. Furthermore, in the case of interest in their appearance image, the point was 3.72, overall, a little above the average. Besides, the majority(51.1%) were not satisfied with their appearance. Besides, the majority(51.1%) were not satisfied with their appearance. As believed by the research, for the solution to the part of the appearance image, body shape management was the most preferred(25.4%), followed by leaving as it is(22.5%) and makeup(20.3%). Moreover, for the monthly average expenditure, 100 thousand won was the most frequent with 26.1%. For the hair style in the present, bobbed hair was topped the list with 20.7%, followed by wave with short cut(20.3%), shoulder-length wave(17.0%), short cut(12.7%). In addition, the degree to which the first impression is affected was 4.35 points on average, very high. On the part of hair style, the degree representing personality was 3.78 points on average, which is slightly high. The degree to show personal expression and aesthetic sense was 4.27 points on average, which is significantly high. Among the psychological factors to decide hair styles, ‘covering aesthetic defects of head shape’ was the most frequent with 42.8%. On the part of hair style, the degree reflecting the culture and thought of the time was 4.11 points on average, which is pretty high. Also, when it comes to changing hair styles, ‘their own judgment’ was the highest with 50.7%

세미퍼머넌트 메이크업 선호도 및 법적규제 인지정도 연구

우정희 ( Jeong-hee Woo ),이근광 ( Keun-kwang Lee ),송연숙 ( Yeon-sook Song ) 대한미용문화예술학회 2017 대한미용문화예술학회지 Vol.6 No.2

To figure out the perception of and satisfaction with cosmetic behavior in semi-permanent makeup, a questionnaire survey was conducted against a total of 1,044 people in their 20-50s living in capital area(Seoul, Gyeonggi and Incheon), Chungcheong area(Daejeon, Cheongju and Cheonan), Honam area(Jeonju, Gwangju and Iksan) and Yeongnam area(Daegu, Ulsan and Busan) from March 30, 2015 to April 30, 2015. The study results found the followings: when asked why they prefer semi-permanent makeup, it is not messed up even during exercise is most responded. when asked they intend to take this treatment again, more than a few respondents said Yes it appeared that many respondents weren’t aware of legal regulations on semi-permanent makeup. When asked if they agree to the legalization of the treatment, ‘I agree’ was mostly responded. Therefore, the said results reveal that semi-permanent makeup has become more common among the general public as well as cosmetologists. In addition, most people wanted to get the treatment at a professional makeup salon, not in a medical institution. In addition, even though many people weren’t aware of legal restrictions on semi-permanent makeup, many of them agree with their legalization. Hence, it is expected that semi-permanent makeup market would further grow. Moreover, since many customers prefer getting the treatment in a professional makeup salon, it is urgent to provide professional education and distribute high-quality skills. Because of a lack of previous studies on semi-permanent makeup, it’s been difficult to get sufficient data on it. In addition, there are legal restrictions in providing this treatment. Therefore, it is hoped that beauty professionals would be able to freely provide this semi-permanent makeup soon. Furthermore, it is essential to establish a license system which can verify semi-permanent makeup under the related laws just like the U.S or through a state exam.