• 한국식품영양학회지
• /
• 제8권4호
• /
• pp.307-317
• /
• 1995

The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

• Bulletin of the Korean Chemical Society
• /
• 제28권12호
• /
• pp.2303-2309
• /
• 2007

Expressed protein ligation (EPL) technique, joining recombinantly expressed proteins to polypeptides, has been widely adopted for addressing various biological questions and for drug discovery. However, joining two recombinant proteins together is sometimes difficult when proteins are expressed insoluble and unrefoldable, because ligation-active proteins via intein-fusion are obtainable when they are folded correctly. We overcame this limitation coexpressing target protein with additional methionine aminopeptidase (MAP) which enhances removal of the initiation methionine of recombinantly expressed protein. Our approach demonstrated that two domains of 46 kDa 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase, a target of herbicide glyphosate, were successfully joined by native chemical ligation, although its C-terminal domain was expressed as an inclusion body. The intein-fused N-terminal fragment of EPSP synthase (EPSPSN, residues 1-237) was expressed and the ligation-active thioester tagged N-terminal fragment (EPSPSN-thioester) was purified using a chitin affinity chromatography and mercapto-ethanesulphonate (MESNA) as intein thiolysis reagent. Its Cterminal fragment (EPSPSC, residues Met237-238CYS-427), expressed as an inclusion body, was prepared from an additional MAP-expressing strain. Protein ligation was initiated by mixing ~1 mM of EPSPSN-thioester with ~2 mM of EPSPSCCYS (residues 238CYS-427). Also we found that addition of 2% thiophenol increased the ligation efficiency via thiol exchange. The ligation efficiency was ~85%. The ligated full-length EPSP synthase was dissolved in 6 M GdHCl and refolded. Circular dichroism (CD) and enzyme activity assay of the purified protein showed that the ligated enzyme has distinct secondary structure and ~115% specific activity compared to those of wild-type EPSP synthase. This work demonstrates rare example of EPL between two recombinantly expressed proteins and also provides hands-on protein engineering protocol for large proteins.

• Clinics in Shoulder and Elbow
• /
• 제8권2호
• /
• pp.63-67
• /
• 2005

이상의 보고에서 임상의 들은 청소년기 운동선수에서 주관절 동통이 있을 때 반드시 골연골염에 대한 주의가 필요하며 비록 치료방법 및 예후에 있어 아직도 많은 논쟁은 있으나 초기발견이 매우 중요한 예후결정의 요소가 된다는 것을 주지하여야 한다. 많은 경우에 있어 초기에 보존적 치료시 증상의 호전 및 기능의 회복 그리고 운동으로의 복귀가 가능하다. 그러나 보존적 치료로 호전이 되지 않을 때 수술적 치료를 고려해야 하는데 이 때 수술적 치료 선택에 매우 신중해야 하며 수술적 적응증 역시 반드시 숙지하여야 한다.

• 미생물학회지
• /
• 제37권4호
• /
• pp.245-252
• /
• 2001

MLS (macrolide-lincosamide-streptogramin B) 항생제 내성인자 단백질인 Erm 단백질들은 아미노산 서열 중 그 동일성과 유사성이 높아 구조적으로도 동등한 단백질의 한 집단을 형성한다. 최근 X-ray crystallography에 의해 구조가 결정된 ErmC\` 및 ErmAM 단백질의 구조에 근거하여 ErmSF 단백질도 catalytic domain과 substrate binding domain으로 구분하였고 N-terminal end에 존재하는 catalytic domain의 대량생산을 다양한 pET 발현 vector를 사용하여 시도하였다. 그리고 catalytic domain을 coding하는 DNA 절편은 세 종류를 사용하였다: DNA 절편 1은 Met 1부터 Glu 186까지를 coding하고 DNA 절편 2는 Arg 60부터 Glu 186까지의 정보를 가진 DNA이고 DNA 절편 3은 Arg 60부터 Arg 240까지를 encoding하는 DNA이다. 사용된 다양한 발현 vector중에서 pET19b는 DNA 절편 3, pET23b는 DNA 절편 1과 2를 성공적으로 대량생산하였다. 그러나 대량생산된 catalytic domain들은 불용성 단백질 집합체인 inclusion body를 형성하였다. ErmSF catalytic domain들의 용해성 단백질의 생산을 위하여 chaperone GroESL과 Thioredoxin의 동시 발현 및 배양온도를 $22^{\circ}C$로 낮추어 시도했으나 대량 발현된 단백질의 용해에는 도움을 얻지 못하였다.

• 미생물학회지
• /
• 제34권3호
• /
• pp.132-136
• /
• 1998

자연환경의 미생물군집을 분자생태학적 방법으로 분석하기 위해서는 오염되지 않은 순수한 핵산의 분리가 필요하다. 해양퇴적물에서 핵산을 추출할 때 같이 추출되는 부식물질(humic substances)은 핵산중폭반응을 저해하기 때문에, 이를 제거하기 위한 네 가지의 방법, 아가로즈젤 전기영동후 용출, Sephadex G-75 젤, Hydroxyapatite 젤, 그리고 PVPP(poluvinylpolypyrrolidone) microspin column 방법에 대해 그 효율성을 비교하였다. 조산된 방법 중에서 PVPP microspin column을 이용하면 건조 퇴적물 1g당 $4.8{\mu}g$의 순수한 DNA를 신속하고 간편하게 얻을 수 있었고, 이 방법으로 분리된 DNA를 PCR의 주형으로 사용한 결과 16S rRNA 유전자의 거의 전 범위에 해당하는 1.5 kb 크기의 PCR 산물을 얻을 수 있었다.

• Journal of Korean Neurosurgical Society
• /
• 제53권3호
• /
• pp.180-182
• /
• 2013

We describe a case of intracranial carotid artery occlusion due to penetrating craniofacial injury by high velocity foreign body that was relieved by decompressive surgery. A 46-year-old man presented with a penetrating wound to his face. A piece of an electric angular grinder disc became lodged in the anterior skull base. Computed tomography revealed that the disc had penetrated the unilateral paraclinoid and suprasellar areas without flow of the intracranial carotid artery on the lesion side. The cavernous sinus was also compromised. Removal of the anterior clinoid process reopened the carotid blood flow, and the injection of glue into the cavernous sinus restored complete hemostasis during extraction of the fragment from the face. Digital subtraction angiography revealed complete recanalization of the carotid artery without any evidence of dissection. Accurate diagnosis regarding the extent of the compromised structures and urgent decompressive surgery with adequate hemostasis minimized the severity of penetrating damage in our patient.

• 대한족부족관절학회지
• /
• 제4권1호
• /
• pp.35-38
• /
• 2000

Osteochondral fracture is common in adolescent and usually occurs in the knee and ankle. Many authors have written concerning osteochondral fracture of the ankle joint, however, many of these pertain only to the talus. The author has found an osteochondral fracture of the articular surface of the medial malleolus within the ankle joint not previously described. We report a case of this lesion that was treated by arthroscopic osteochondral fragment removal.

• Journal of Trauma and Injury
• /
• 제32권2호
• /
• pp.122-125
• /
• 2019

A 73-year-old man, who, in an inebriated state, had slipped in a flowerbed and was wounded on the left flank, was transferred to Trauma Center, Gil Medical Center, Gachon University College of Medicine. Based on the chest and abdominopelvic computed tomography, he was diagnosed with multiple rib fractures and hemopneumothorax on the left hemithorax and was found to have a bony fragment in the spleen. He had not presented peritonitis and exsanguinous symptoms during the observation period. Seven days later, computed tomography of the abdomen showed suspected diaphragmatic injury and a retained foreign body in the spleen. On exploration by video assisted thoracoc surgery (VATS), a herniated omentum through the lacerated site of the diaphragm was observed. After omentectomy using Endo Gia, the foreign body in the spleen was observed through the lacerated site of the diaphragm. Traumatic diaphragm rupture with a foreign body, in the spleen, was successfully managed by video assisted thoracic surgery via the lacerated site of the diaphragm.

• Journal of Dental Anesthesia and Pain Medicine
• /
• 제22권4호
• /
• pp.315-321
• /
• 2022

Although rare, hypodermic needle fractures can occur in the maxillofacial region. In cases of fracture, urgent intervention is required to prevent further complications. We present the case of a 37-year-old female patient with a fractured needle in the left sublingual fossa during a lingual nerve block 6 months before referral. The fragment of a 30-gauge needle was located using cone-beam computed tomography and retrieved under local anesthesia with blunt dissection. The patient recovered uneventfully, except for predictable postoperative inflammatory complications, which resolved within 2 weeks. Precautions should be implemented to prevent needle fractures, which are usually preventable. However, if the retrieval is unsuccessful, the patient should be referred to a well-equipped surgical unit without delay.

• 한국물환경학회지
• /
• 제36권3호
• /
• pp.220-228
• /
• 2020

Anaerobic ammonium oxidation (AMX) is a cost-efficient biological nitrogen removal process. The coexistence of ammonia-oxidizing bacteria (AOB) in an AMX reactor is an interesting research topic as a nitrogen-related bacterial consortium. In this study, a sequencing batch reactor for AMX (AMX-SBR) was operated with a conventional activated sludge. The AOB in an AMX bioreactor were identified and quantified using terminal restriction fragment length polymorphism (T-RFLP) and real-time qPCR. A T-RFLP assay based on the ammonia monooxygenase subunit A (amoA) gene sequences showed the presence of Nitrosomonas europaea-like AOB in the AMX-SBR. A phylogenetic tree based on the sequenced amoA gene showed that AOB were affiliated with the Nitrosomonas europaea/mobilis cluster. Throughout the enrichment period, the AOB population was stable with predominant Nitrosomonas europaea-like AOB. Two OTUs of amoA_SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments. Real-time qPCR was used to quantify AOB populations over time. Interestingly, the exponential growth of AOB populations was observed during the substrate inhibition of the AMX bacteria. The specific growth rate of AOB under anaerobic conditions was only 0.111 d^-1. The growth property of Nitrosomonas europaea-like AOB may provide fundamental information about the metabolic relationship between the AMX bacteria and AOB.