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Blunt End Ligation 의 최적반응에 관한 연구
송옥규,김용석,노현모 ( Ok Kyu Song,Yong Sok Kim,Hyune Mo Rho ) 생화학분자생물학회 1985 BMB Reports Vol.18 No.3
To determine the optimal condition of blunt-ended DNA ligation for the purpose of the efficient cloning, blunt-ended vector and insert DNA fragments were ligated under the various conditions of buffer, temperature, reaction time, and the molar ratio of vector to insert DNA. The efficiency of ligation was monitored by both agarose gel electrophoresis and the transforming frequency of genetic markers in plasmid pRSK 116 in vivo. The results showed that the optimal incubation temperature and the concentration of ATP were 20-25℃ and 0.5 mM, respectively. Spermidine (1 mM) and polyethylene glycol (3%) stimulated blunt-end ligation, but more than 10 mM of spermidine and 2 mM of ATP completely inhibited the reaction. Higher concentration of salt also inhibited blunt-end ligation. According to the transforming frequency assay, the optimal molar ratio of vector to insert DNA was 1, which was different from that of sticky-end ligation. However, the variation of pH (6.6-8.2) and bovine serum albumin concentration (0-500 ㎍/㎖) showed little influence on the ligation.
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Studies on the Optimal Condition for Ligation of Blunt Ended DNA Fragments
송옥규,김용석,노현모,Song, Ok-Kyu,Kim, Yong-Sok,Rho, Hyune-Mo 생화학분자생물학회 1985 한국생화학회지 Vol.18 No.3
Blunt end DNA ligation의 최적반응조건을 구하기 위하여 반응온도, 완충용액상태, vector와 insert DNA의 농도비율, 반응시간등의 최적조건을 연구하였다. 실험결과 반응최적온도는 $20-25^{\circ}C$였고, ATP의 최적농도는 0.5 mM이었다. Spermidine의 농도가 1 mM 인 경우와 4%의 PEG 4,000을 ligation buffer에 넣어준 경 우 반응이 잘 되는 것으로 나타났으나 spermidine이 10 mM이거나 ATP의 농도가 2 mM 이상에서 반응은 저해되었다. 높은 농도의 NaCl 또한 반응을 저해시키는 것으로 나타났다. Vector와 insert DNA의 농도비를 달리하여 ligation시켜 이것을 직접 E. coli를 형질 전환시켜 본 결과 농도비가 1일때 insert가 끼어 들어간 확율이 가장 높았다. Buffer의 pH변화(6.6-8.2)나 BSA의 농도변화 ($0-500\;{\mu}g/ml$)는 반응에 별 영향이 미치지 않는 것으로 나타났다. To determine the optimal condition of blunt-ended DNA ligation for the purpose of the efficient cloning, blunt-ended vector and insert DNA fragments were ligated under the various conditions of buffer, temperature, reaction time, and the molar ratio of vector to insert DNA. The efficiency of ligation was monitored by both agarose gel electrophoresis and the transforming frequency of genetic markers in plasmid pRSK 116 in vivo. The results showed that the optimal incubation temperature and the concentration of ATP were $20-25^{\circ}C$ and 0.5 mM, respectively. Spermidine (1 mM) and polyethylene glycol (3%) stimulated blunt-end ligation, but more than 10 mM of spermidine and 2 mM of ATP completely inhibited the reaction. Higher concentration of salt also inhibited blunt-end ligation. According to the transforming frequency assay, the optimal molar ratio of vector to insert DNA was 1, which was different from that of sticky-end ligation. However, the variation of pH (6.6-8.2) and bovine serum albumin concentration ($0-500\;{\mu}g/ml$) showed little influence on the ligation.
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Blunt - end 를 가진 DNA 의 재조합을 위한 유전자 운반체의 개발
노현모,김용석,송옥규 한국유전학회 1984 Genes & Genomics Vol.6 No.2
A blunt-end DNA cloning vector, pRSK 116, was constructed. This vector was derived from pBR 322 by opening with Sal I restriction endonuclease, filled with only TTP by Klenow fragment, trimmed with S1 unclease, and ligated with T4 DNA ligase. It appeared that the removal of three bases in the middle region of Tc^r gene did not change the genetic property of the marker gene of the pBR 322 and the unique Hinc II site in the Ap^r gene of the modified plasmid pRSK 116 en. dowed improved property for the selection of recombinants. The Hinc II site has been tested for the cloning of blunt-ended human DNA. The recombinant colonies selected were Ap sensitive and Tc resistant and inserted DNAs were easily recovered by Hinc II treatment.
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Construction of pRSK116 and Modified Method for Blunt - end Ligation of HincⅡ Site of the Plasmid
노현모,김용석,송옥규 한국유전학회 1983 Genes & Genomics Vol.5 No.3
We have constructed plasmid pRSK116 which has only one site of HincII restriction endonuclease within the β-lactamase gene which is resistant to ampicillin. The pRSK116 was derived from pBR322 by the following procedures. Plasmid pBR322 was opened by treatment with SalI endonuclease, filled with only TTP by Klenow fragment, trimmed with S1 nuclease, and ligated with T4 ligase. We have used the HincII site of the plasmid, pRSK116, for the study of blunt-end ligation since the pBR322 does not have blunt-end restriction site within marker genes. We attempted to ligate blunt-end DNA fragments with T4 DNA ligase under various conditions (e.g. temperature, DNA concentration, and incubation time), and measured the ligation efficiency by agarose-gel electrophoresis and transformation in E.coli HB101 in the presence of proper antibiotics.
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이홍선,이영한,정인하,송옥규,김성준,송호택,서진석 대한영상의학회 2020 대한영상의학회지 Vol.81 No.1
Magnetic resonance imaging (MRI) is an essential modality for the diagnosis of musculoskeletal system defects because of its higher soft-tissue contrast and spatial resolution. With the recent development of MRI-related technology, faster imaging and various image plane reconstructions are possible, enabling better assessment of three-dimensional musculoskeletal anatomy and lesions. Furthermore, the image quality, diagnostic accuracy, and acquisition time depend on the MRI protocol used. Moreover, the protocol affects the efficiency of the MRI scanner. Therefore, it is important for a radiologist to optimize the MRI protocol. In this review, we will provide guidance on patient positioning; selection of the radiofrequency coil, pulse sequences, and imaging planes; and control of MRI parameters to help optimize the MRI protocol for the six major joints of the musculoskeletal system. 자기공명영상(magnetic resonance imaging; 이하 MRI)은 다른 영상 기법에 비해 연부 조직 대조도와 해상력이 높아 근골격계 영역에서 중요한 진단 기기로 이용되고 있다. 최근MRI 관련 기술이 발달함에 따라 빠른 영상 촬영 및 다양한 영상면 재구성이 가능해짐으로써입체적인 근골격계 해부학적 구조와 병변을 더욱 잘 평가할 수 있게 되었다. 또한, MRI는 최적화 정도에 따라 영상의 질, 진단 정확도 및 촬영 시간 등이 달라지며, MRI 장치의 효율적인 운용과도 관련이 있어, 이를 관리하는 것은 영상의학과 의사의 중요한 역할이다. 본 종설에서는 6개 주요 관절에 따른 환자 자세, radiofrequency 코일 선택, 권장 펄스열, 영상면구성 및 스캔 파라미터에 대한 지침을 제시함으로써 근골격계 MRI의 최적화에 도움이 되고자 한다.
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