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Jang, Sei-Heon,Lee, Chang-Hwan The Korea Science and Technology Center 1998 BMB Reports Vol.31 No.6
Transcription of mitochondrial DNA in the yeast S. cerevisiae depends on recognition of a consensus nonanucleotide promoter sequence by mitochondrial RNA polymerase specificity factor, which is a 43 kDa polypeptide encoded by the nuclear MTF1 gene. Mtf1p has only limited amino acid sequence homology to bacterial sigma factors, but functions in many way like sigma in that it is required for promoter recognition and initiation of transcription. To analyze the core-binding region of Mtf1p, monoclonal antibodies to this protein were prepared. Recombinant Mtf1p over-produced in E. coli was purified to near homogeneity and used to raise monoclonal antibodies (mAbs). From fused cells screened for Mtf1p mAbs by immunodot blot analysis, 19 positive clones were initially isolated. Further analysis of positive clones by Western blotting resulted in 4 mAbs of Mtf1p.
Construction and purification of yeast recombinant Rpo41p core polymerase
Jang, Sei-Heon 대구대학교 기초과학연구소 2003 基礎科學硏究 Vol.19 No.3
Yeast mitochondrial RNA polymerase is a good model system for studying eukaryotic gene transcription because it simply consists of two subunits and recognizes promoter sequences similar to that of nuclear RNA polymerase. To promote work on transcriptional mechanism of mitochondrial RNA polymerase, recombinant form of RPO41 core polymerase was constructed, over-expressed in E. coli and purified to near homogeneity using a Ni+-NTA column and conventional biochemical columns for crystallization. On the basis of protein yields and specific activity of recombinant Rpo41p, we estimated that active recombinant Rpo41p can be purified at the yield rate of 1 ㎎ per 1 g of E. coli cells induced for the production of Rpo41p by 0.6 mM IPTG.
Amino acid substitutions conferring cold-sensitive phenotype on the yeast MTF1 gene
Jang, Sei-Heon The Microbiological Society of Korea 1997 The journal of microbiology Vol.35 No.3
The MTF1 gene of Saccharomyces cerevisiae encodes a 43 kDa MITOCHONDRIAL RNA polymerase specificity factor which recognizes mitochondrial promoters to initiate correct transcription. To better understand structure-function of the MTF1 gene as well as the transcription mechanism of mitochondrial RNA polymerase, two cold-sensitive alleles of the MTF1 mutation were isolated by plasmid shuffling method after PCR-based random mutagenesis of the MTF1 gene. The mutation sites were analyzed by nucleotide sequencing. These cs phenotype mtf1 mutants were respiration competent on the nonfermentible glycerol medium at the permissive temperature, but incompetent at 13.deg.C. The cs phenotype allele of the MTF1, yJH147, encoded an L146P replacement. The other cs allele, yJH148, contained K179E and K214M double replacements. Mutations in both alleles were in a region of Mtflp which is located between domains with amino acid sequence similarities to conserved regions 2 and 3 of bacterial s factors.
Sung-Min Lee,Do Kyung Hong,Sei-Heon Jang,ChangWoo Lee 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10
The longhorn pine sawyers, Monochamus saltuarius and M. alternatus (Coleoptera: Cerambycidae), are vectors of the pinewood nematode Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae), the causal pathogens of pine wilt disease in Korea. Recently, an aggregation pheromone, 2-(undecyloxy)ethanol, identified from M. galloprovincialis and M. alternatus, was shown to be effective for attracting several Monochamus species in Europe, North America, and East Asia. However, the effect of 2-(undecyloxy)ethanol on attracting M. saltuarius remains largely unraveled. In this study, we investigated the abilities of 2-(undecyloxy)ethanol along with host plant volatiles (α-pinene and ethanol) to attract M. saltuarius at a pine forest in Cheongsong, Gyeongsangbuk-do, Korea. Pine trees in the field experiment site were not previously affected by pine wilt disease. The combination of 2-(undecyloxy)ethanol with host plant volatiles was more effective than either of 2-(undecyloxy)ethanol or host plant volatiles for attracting M. saltuarius. Both sexes of M. saltuarius were attracted to traps containing 2-(undecyloxy)ethanol with the host plant volatiles. Our study suggests that the aggregation pheromone in combination with host plant volatiles could be used for detection and population monitoring of M. saltuarius as well as for effective mass trapping in outbreak situations.
The experience of transumbilical endoscopic appendectomies
Chung-Heon Lee,Won Joong Jeon,Sei Jin Youn,Hyo Young Yun,Lee-Chan Jang,Jae-Woon Choi,Young Jin Song,Dong Hee Ryu 대한외과학회 2014 Annals of Surgical Treatment and Research(ASRT) Vol.86 No.5
Minimally invasive surgery is being widely accepted in various fields of surgery. Although several appendectomy techniques have been reported but, there is no standardization. We report here the experiences of transumbilical endoscopic appendectomy in humans. Between July 2008 and September 2010, ten patients with appendicitis successfully underwent transumbilical endoscopic appendectomies. There were 7 cases of suppurative, 2 cases of gangrenous and 1 case of perforated in operative findings. The ages of the patients were 13?56 years (mean age, 32.7 ± 15.4 years). Under general anesthesia, a 15-mm port was inserted through the umbilicus and then a two-channel endoscope was inserted in the peritoneal cavity. After appendix identification, counter-traction of the appendix with a direct abdominal wall puncture using a straight round needle prolene was performed to achieve good visualization of the operative field. Tissue dissection was performed using an endoscopic needle knife. Tissue grasping and resected appendix retrieval were done with endoscopic forceps. The average operation time was 79.5 ± 23.6 minutes (range, 45 to 110 minutes). No procedures were converted to laparoscopic or open appendectomy. Hospital stay was 4?6 days. All patients completely recovered without complications. As it is highly maneuverable, we believe transumbilical endoscopic appendectomy can be a feasible method. And, as surgeons want to proceed from laparoscopic surgery to natural orifice transluminal endoscopic surgery, this procedure could be a triable method.