Agrobacterium tumefaciens의 Ti plasmid 재조합에 관한 연구

주충노,이희봉,김영명 연세대학교 자연과학연구소 1987 學術論文集 Vol.19 No.-

Crown gall의 병원체로 알려진 Agrobacterium tumefaciens가 함유하고 있는 Tumor inducing(Ti) plasmid가 식물 hormone과 opine의 합성원인이라는 것이 밝혀지고 식물 세포의 crown gall에는 Ti plasmid가 존재하며 Ti plasmid의 일부가 식물세포의 핵 genome에 삽입되어 형질 변화가 일어남이 보고 되었고 Ti plasmid의 식물체로의 gene도입 vector로서의 연구가 최근 활발히 진행되고 있으며 현재 PLGV vector와 PMON vector 등이 개발되고 있으나 조작이 복잡하고 아직은 초보적 단계인 것으로 알려져 있다. 본 연구에서는 원하는 식물 gene을 식물체에 도입하는 유용한 vector개발의 초보단계로서 Ti plasmid의 T-DNA의 일부인 nos gene을 분리하여 pBR322와의 재조합을 기도한 것이다. pBR 322를 포함하는 E. coli로 부터 Horwitz(1979)의 방법으로 pBR 322를 분리하였고 A. tumefaciens C58에서 Kado(1979)의 방법으로 Ti plasmid를 분리한 후 Ti plasmid 내의 nos gene을 분리하였다. 분리된 pBR 322를 Hind Ⅲ로 절단한 후 CIP로 처리하고 T_4 DNA ligase를 이용하여 Hind Ⅲ로 처리한 nos gene fragment와 재조합하였다. 이와같이 하여 얻은 recombinant DNA로 인한 E. coli HB101의 형질 변화를 Mandel과 Higa의 방법에 따라 조사한 결과 pBR 322의 tetracyclin gene 사이에 nos gene이 삽입된 것으로 확인되었다. 즉 ampicillin 배지에서는 92개의 colony가 형성되었는데 그중 59개는 tetracyclin 배지에서 colony를 형성하지 못하였다. 이러한 tetracyclin 감수성이면서 ampicillin 저항성인 colony를 배양하여 얻은 recombinant plasmid를 전기이동법으로 분리한 결과 예상대로 크기가 7.5kb 정도였고 Hind Ⅲ로 절단하였을 때 2개의 band가 확인되었다. 본 연구에서 얻은 nos gene-pBR 322 hybrid plasmid는 원하는 식물 gene을 이것에 삽입하여 A. tumefaciens 내에서 homologous recombination을 행한 후 식물체에 감염시켜 원하는 gene의 발현여부를 검출하는데 이용될 것으로 기대된다. It is mow realized that the tumor-inducing(Ti) plasmid in Agrobacterium tumefaciens is responsible for the induction of crown gall tumors in dicotyledonous plants and it has been demonstrated that a segment of the Ti plasmid, so called T-DNA, is stably integrated into and expressed in the genome of transformed plant cells. Recently, the use of A. tumefaciens Ti plasmid as a vector to introduce a foreign gene has been intensively studied and several vectors such as PLGV vector and PMON vector has been reported. However, their practical use is still at a primitive stage. It was attempted in the present study, therefore, to conduct the recombination of nos gene of T-DNA from A. tumefaciens Ti plasmid and E. coli pBR 322 as a vector it introduce foreign plant gene into dicotyledon plant cells. pBR 322 was isolated from E. coli containing pBR 322 accdrding to Horowitz(1979) and nos gene of T-DNA in Ti plasmid from A. tumefaciens C58 was obtained according to Kado(1979). The isolated pBR 322 was cut using Hind Ⅲ followed by the addition of CIP·pBR 322 fragments and Hind Ⅲ treated nos gene fragments were than recombined using T_4 DNA ligase and the E. coli cells were then transformed by recombinant DNA according to Mandel and Higa(1970). It was confirmed that nos gene was inserted into tetracyclin gene of pBR 322 by the finding that when the transformed cells were incubated in the ampicillin medium, 92 colonies were formed, in which 59 were tetracyclin sensitive. The above tetracyclin sensitive but ampicillin resistant cells were then cultured and the recombinant pBR-nos gene was isolated and subjected to electrophoresis. It was found that size of the recombinant DNA was approximately 7.5 Kb as expected and two bands were appeared on electrophoretogram when the recombinant DNA was treated with Hind Ⅲ. It might be possible to insert the foreign plant genes to the pBR-nos gene hybrid plasmid obtained in the present study and the resultant DNA could be then homolgously recombined with Ti plasmid of A. tumefaciens and the final recombined Ti plasmid could be used for the introduction of the foreign plant genes into dicotyledon plant cells.

중심압축력을 받는 高强度 RC기둥의 띠철근 配筋形態에 關한 實驗的 硏究

이영인,이영호,곽노현,정헌수 중앙대학교 기술과학연구소 1998 기술과학연구소 논문집 Vol.28 No.-

An objective of this study is to experimentally investigate the strength and ductility of reinforced concrete columns under uniaxial load and several test variables. To do this, we have conducted tests on twelve 20×20×60cm specimens with 8 and 12 longitudinal steel bars subjected to monotonic uniaxial compression. The main variables considered in this test are the configuration of ties, the strength of concrete, and the space of ties. The results indicate that the strength and the ductility of reinforced concrete columns have been influenced on the configuration of ties, the space of ties, and the strength of concrete.

中心壓軸力을 받는 高强度 鐵筋콘크리트 기둥의 耐力과 延性에 關한 실험적 硏究

곽노현,이영호,김남주,이경종,정헌수 중앙대학교 기술과학연구소 1997 기술과학연구소 논문집 Vol.27 No.-

A study on the behavior of short reinforced concrete columns was experimently described. Twenty-four concrete columns, with 20cm square and 60cm height, containing 8 longitudinal steel bars and different the amount of ties, spacing and concrete strengths, were subjected to concentric load by failure. Test results indicated that 1) It is incompatible in high-strength concrete to arrange stirrups to the AIK code of D/2 throughout the example of 10cm stirrup spacing following AIK codes. 2) As the amount of stirrup increased, the strength and ductily of reinforced concrete column improved. 3) As the concrete strength is high, the effect of the was reduced. 4) If strength enhancement factors increased, strain at maximum load also does and ductility ratios does almost linearliary. 5) The rate of strength enhancement factor does well with the modified kent & Park's theory. 6) As to the result of calculating the effective confined area which is suggested in Sheikh & Uzumeri's theory, the middle vertical steel is effective, only if the stirrups are arranged with spacing of 3cm.

균 spheroplast 및 large unilamellar vesicle의 식물 protoplast로의 도입

주충노,강철호,이희봉 연세대학교 자연과학연구소 1987 學術論文集 Vol.19 No.-

A. tumefaciens의 식물 protoplast로의 도입을 확인하는 방법으로 방사성 인산(Na_2H^32PO_4)을 함유한 0.8% nutrient broth에서 증식한 Agrobacterium tumefaciens C58를 사용하여 방사성 spheroplast와 방사성 Ti plasmid encapsulated liposome을 만들어 Danus carota protoplast로의 도입을 시도한 결과 다음과 같은 실험 성적을 얻었다. 1. A. tumefaciens에서 추출한 Ti plasmid를 Reverse Phase Evaporation 법으로 phosphatidyl-choline, stearylamine과 함께 Ti plasmid encapsulated liposome을 제조한 결과 17%의 Ti plasmid가 liposome안으로 편입되었다. 2. 방사성 Ti plasmid encapsulated liposome과 Dacus carota root protoplast를 polyethyleneglycol-Ca^2+법으로 처리한 결과 introduction frequency는 38%이었다. 3. A. tumefaciens C58의 spheroplast와 Dacus carota root protoplast를 polyethyleneglycol Ca^2+ 법으로 처리한 결과 introduction frequency가 47.6%였다. 4. 본 연구에서 얻은 실험결과로 부터 liposome법, spheroplast법 모두 Ti plasmid의 식물 protoplast로의 도입은 용이하게 일어나는 것으로 생각된다. It was attempted in the present study to prepare radioactive Ti plasmid to identify the introduction frequency of Ti plasmid encapsulated liposome into Dacus carota root protoplast. Agrobcterium tumefaciens C58 was cultured in 0.8% nutrient broth containing radioactive phosphoric acid(H_3^32PO_4) and the radioactive Ti plasmid wad isolated. Radioactive Ti plasmid was encapsulated into phosphatidyl choline-stearylamine liposomes by reverse phase evaporation technique and the encapsulation frequency was found to be about 17%. Radioactive Ti plasmid encapsulated liposome and Dacus carota root protoplast was treated by polyethyleneglycol Ca^2+ procedure. The introduction frequency was 38%. The spheroplast of A. tumefaciens C38 was prepared from the bacteria cultured in 0.8% nutrient broth containing radioactive phosphoric acid using lysozyme. As much as 47% of the spheroplast were found to be introduced into Dacus carota protoplast.

구속된 고강도 콘크리트 기둥의 휨 거동에 관한 실험적 연구

양근혁,이영호,곽노현,정헌수 중앙대학교 기술과학연구소 1998 기술과학연구소 논문집 Vol.28 No.-

본 실험적 연구의 목적은 콘크리트 강도, 띠근 배근형태, 띠근 량, 축하중비가 축하증과 횡하증을 받는 구속된 콘크리트 기둥의 휨 내력과 연성에 미치는 영향을 팡가하기 위한 것이다. 구속된 기둥의 휨 거동을 시험하기 위하여 200×200×800mm 크기의 시험체 14개를 2축 하중하에서 실험하였다. 시험결과로서, 저축력에서 콘크리트 구속으로 인한 휨 강도의 증가는 비교적 작게 있지만, 고축력에서 콘크리트 구속으로 인한 휨 강도 증가는 크게 있다. 그리고, 고축력, 고강도 콘크리트에서 내부 cross-ties가 있으면 휨 연성의 증가가 보였으며, 구속된 콘크리트 기둥의 연성은 단순한 띠근 량보다는 구속지수에 관계되었다. The purpose of this experimental study was to investigate the effects of concrete strength, the configuration of lateral ties, the amount of lateral ties, and level of axial loads on flexural strength and ductility of confined of reinforced concrete columns subjected to combined axial and lateral loads. To simulate the flexural behavior of confined columns, fourteen reinforced concrete columns, 200×200×800mm, were tested under bi-axial loads. Test results indicate that for low axial load levels, the enhancement of the flexural strength due to concrete confinement is relatively small, but that for high axial load levels, the enhancement of the flexural strength due to concrete confinement can be very high. Considerable enhancement of ductility was observed in columns confined by cross-ties, even for high strength concrete columns and under high axial loads. Ductility of confined columns are found to be more related to confinement index (ρ??f??/??) than the amount of lateral ties(ρ??)

Al-Zn-Mg-Cu-Zr 합금의 집합조직 발달과 기계적 특성에 미치는 Sc 첨가의 영향

제창웅,진승준,정동석,임성택,이용연,박노진 대한금속재료학회 2004 대한금속·재료학회지 Vol.42 No.10

The texture and mechanical properties during fabrication processes - extrusion, cold rolling and aging treatment - of plate in the 7050 Al alloys without/with 0.1%Sc were investigated. In both the cross section(CS) and longitudinal section(LS) of the specimen without Sc, after cold rolling, texture of R-fiber was predominant. In the Sc added alloy after cold rolling, {112}<111>(Cu)+weak (123)<634>(S) texture components developed in the CS of the specimen while strong (3-fiber+(110)<001>(Goss) components developed in the LS of the specimen. After solution treatment and aging treatment, the alloy (CS and LS) without Sc had random texture, but by the alloys with the 0.1%Sc, the components of the rolling texture are not changed. After the aging treatment, hardness and tensile strength of the alloy with 0.1%Sc increased about 15% and 17%, respectively, than those without Sc. Tensile strength of the Sc added specimen was the highest along the rolling direction where the <111> texture was strongly developed. On the other hand, in the specimen without Sc, the mechanical properties were not dependent on the direction of the sample. (Received May 3, 2004)

電氣工學敎育의 發展方向에 關한 硏究

李鎭局,徐相壽,朴魯峰,李忠鎬,劉鍾哲,金洪奎,趙武濟,韓運東,池平植 충주대 2003 産業科學論文集 Vol.11 No.-

In this paper, we discuss actual conditions for running school of electrical and electronic engineering in domestic universities. Also, we catch hold of industrial circles's demands in electrical engineering education and analyze questionary results according to goes into effect school of electrical and electronic & information engineering in major of electrical and electronec engineering's students. Conclusively, we present suitable model in electrical engineering by correction of problems.

생강나무(Lindera obtusiloba Blume) 잎 70% 에탄올 추출물의 단회와 14일 반복투여 독성시험의 안전성 평가

홍충의(Chung-Oui Hong),서문영(Young Seomun),구윤창(Yun-Chang Koo),남미현(Mi-Hyun Nam),이현아(Hyun Ah Lee),김지훈(Ji-Hoon Kim),왕증(Wang-Zeng),양성용(Sung Yong Yang),이성희(Sung-Hee Lee),노수환(Su Hwan No),이광원(Kwang-Won Lee) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.10

생강나무는 한국, 일본, 중국 등지에서 자생하며 열, 복통, 타박상, 출혈 등에 민간요법으로 사용되어온 식물이다. 이 논문의 목적은 오래전부터 민간요법으로 사용되어온 생강나무 잎의 안전성을 평가하고자 생강나무 잎 70% 에탄올 추출물을 암수 각 군당 5마리씩 5000(단회투여 독성시험), 0, 500, 1000, 2000(14일 반복투여 독성시험) ㎎/㎏의 용량으로 단회경구투여와 14일 반복경구투여를 실시하여 15일간의 사망률, 일반증상, 체중변화, 조직병리, 혈액학적 및 혈액 생화학적 검사를 실시하였다. 시험기간 중 사망동물은 암수 모든 시험군에서 관찰되지 않았으며, 일반증상, 체중변화, 조직병리, 혈액학적 및 혈액생화학적 검사에서 투여와 관련된 변화는 관찰되지 않았다. 이상의 결과로 보아 생강나무의 단회투여 독성은 5000 ㎎/㎏ 용량을 상회하며, 14일 반복투여의 최고용량인 2000 ㎎/㎏ 용량에서도 아무런 독성이 나타나지 않는 것으로 보아 최대무독성량은 이 농도를 상회하는 것으로 판단된다. 이로써 생강나무는 안전한 소재로서 식재료 또는 건강기능식품의 원료로서도 사용이 가능하다고 볼 수 있다. Lindera obtusiloba Blume (LO), which is widely distributed in Korea, Japan and China, has traditionally been used as a popular folk medicine for the treatment of fever, abdominal pain, bruise and extravation. The purpose of this study was to examine the toxicities of the single and 14-day repeated doses in Sprague-Dawley rats orally administrated with LO at doses of 0, 500, 1000, 2000 (14-day repeated toxicity test) and 5000 (single toxicity test) mg (dry weight)/㎏ of body weight/day. The results showed that there was no difference in body weight change, food intake, water consumption, or organ weight among different dose groups. Also we observed no death and abnormal clinical signs during the experimental period. Between the groups orally administered LO and the control group, there was no statistical significance in hematological test or serum biochemical values. There were no evidences of histopathological alteration as well as abnormal gross finding mediated by single and 14 days treatments with LO. These results suggest that no observed adverse effect level (NOAEL) of the oral application of LO was considered to be more than 2000 mg/kg in rats under the conditions employed in this study.

임신 중기 태아 맥락총 낭종 119예의 추적 조사: - 18번 삼염색체증과의 연관성 여부를 중심으로 -

이은혜(Eun Hye Lee),이유미(You Me Lee),신명철(Myung Choel Shin),민유선(Yu Seon Min),이상희(Sang Hee Lee),김현철(Hyeon Chul Kim),김종욱(Jong Wook Kim),이숙환(Sook Hwan Lee),이위현(Wee Hyun Lee),조진호(Jin Ho Cho),이정노(Chung No Lee) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.7

Objective: To evaluate the clinical significance of fetal choroid plexus cysts (CPCs) in the second trimester, especially an association with trisomy 18. Methods: From March 1998 through June 1999, second trimester screening ultrasonography was performed on 4,948 unselected single-ton pregnancies. CPCs were noted in 132 fetuses. Among them, detailed ultrasonography and follow-up was possible in 119 cases and they were recruited into the study. There were 91 cases of isolated CPCs and 28 cases of CPCs in high-risk population. Isolated CPCs were defined as: mother did not have any risk factors requiring amniocentesis and there were no other sonographic abnormalities on detailed ultrasound. CPCs in high-risk population were defined as: mother had any risk factor requiring karyotyping or there were any other sonographic abnormalities although she was general population. Amniocentesis was performed in 39 cases. We compared gestational age at time of detection, size, bilaterally, multiplicity, and complexity of CPCs in the group of isolated CPCs and CPCs in high-risk population (t-test, chi-square test; p<0.05). We evaluated the findings of detailed and follow-up ultrasonography, karyotypes, and final outcomes of pregnancy.Results: Gestational age at time of detection was not different in both groups of isolated CPCs and CPCs in high-risk population (19±2 vs 18±1 wk, p>0.05). Mean size (6.4 vs 6.2 mm), bilaterality (60% vs 57%), multiplicity (66% vs 57%), and complexity (8% vs 14%) of CPCs were also similar. All CPCs were disappeared irrespective of size and mean time of disappearance was 25±3 and 26±3 week, respectively (p>0.05). All cases of isolated CPCs resulted in phenotypically-normal neonates. It was confirmed by either amniocentesis or postnatal examination by the pediatrician. Among fetuses having CPCs in high-risk population, two trisomy 18 and one trisomy 21 were detected. All of them had positive result of maternal serum marker test and/or sonographic abnormalities. Remaining cases were proved normal.Conclusion: The risk of chromosome abnormalities is very high when CPCs are associated with other abnormalities on detailed ultrasound, indicating a clear need to offering genetic amniocentesis. As contrast, the risk of chromosome abnormalities for a case of isolated CPCs is very low, and in this series there was no trisomy 18. Therefore isolated CPCs should be considered as the indication of detailed ultrasound examination, but not routine karyotyping.

Characterization of Humanized Antibody Produced by Apoptosis-Resistant CHO Cells under Sodium Butyrate-Induced Condition

( No Soo Kim ),( Kern Hee Chang ),( Bo Sup Chung ),( Sung Hyun Kim ),( Jung Hoe Kim ),( Gyun Min Lee ) 한국미생물생명공학회 2003 Journal of microbiology and biotechnology Vol.13 No.6