이중 in situ hybridization기법을 이용한 흰쥐 뇌의 Vasoactive intestinal polypeptide, Vasopressin 및 Oxytocin mRNA의 발현에 관한 연구

박경한,홍석,유영복,이왕재,황덕호,이병란,차중익,조사선,백상호 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.6

Shiga 독소 유전자를 이용한 Colony Hybridization에 의한 장출혈성 대장균의 동정

신형식 대한감염학회 2002 감염 Vol.34 No.4

전자현미경 In Situ Hybridization에 의한 Viral RNA의 진단에 관한 연구

최원기(Won-Gi Choi),주경웅(Keng-Woong Joo),김석홍(Suk-Hong Kim) 대한의생명과학회 1996 Biomedical Science Letters Vol.2 No.2

토끼 바이러스성 출혈증의 원인체를 실험 토끼에 접종하여 증식을 유도하고 간장에서 hematoxylin & eosin 염색에서 조직학적 진단과 세포내 viral RNA의 소재를 결정하기 위해 post-unicryl 포매한 block의 절편을 사용하여 단 염색과 전자현미경적 in situ hybridization을 시도하였다. 토끼 출혈증 viral RNA의 보합 결합에 이용하는 probe는 4717에서 4800(84bases)까지 oligonucleotide를 5’ 말단에 biotin-CE phosphoramidite로 표지하여 사용하였다. 보합결합물의 증명은 신호 표지로서 antibiotin antibody-10㎚ gold를 사용하였으며, hybridization이나 증명은 기존 protocol에서 약간의 변법을 사용하였다. 0.02% glutaraldehyde에서 고정하고 unicryl resin 포매한 표본, biotinylated oligonucleotide probe, antibiotin antibody-10㎚ gold로 실험한 결과 증강된 선호를 얻을 수 있었다. 특히 전처리를 생략하므로써 실험 과정을 간단하게 하여 신속한 결과를 얻을 수가 있었다. 전자현미경 in situ hybridization을 통하여 토끼 출혈증 바이러스의 주요 표적은 간세포로 감염 세포의 세포질 내 미토콘드리아와 핵사이에서 immuno gold입자가 뚜렷하게 표지됨으로서 viral RNA를 증명할 수 있었다. Simple stain and electron microscopic in situ hybridization is studied and applied for the identification of rabbit haemorrhagic disease viral RNA in a unicrylated preparation of the liver after innoculation of rabbit haemorrhagic disease virus. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probe labelled by biotin CE-phosphoramidite compared with 4717~4800 sequences of rabbit haemorrhagic disease virus, modified hybridization protocol and antibiotin antibody-10㎚ gold as signal marker. The best results were obtained in 0.02% glutaraldehyde, Unicryl resin cell block, biotinylated oligonucleotide probes, antibiotin-10㎚ gold. In this report, RHD viral RNA was distributed widely within the mitochondria and nucleus of liver cell by electron microscopic in situ hybridization. In situ hybridization has become a standard method for localizing DNA or RNA sequences in tissue or cell preparation. In situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method.

전자현미경 In Situ Hybridization에 의한 Viral RNA의 진단에 관한 연구

김석홍,주경웅,최원기 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1996 Journal of biomedical laboratory sciences Vol.2 No.2

토끼 바이러스성 출혈증의 원인체를 실험 토끼에 접종하여 증식을 유도하고 간장에서 hematoxylin & eosin 염색에서 조직학적 진단과 세포내 viral RNA의 소재를 결정하기 위해 post-unicryl 포매한 block의 절편을 사용하여 단 염색과 전자현미경적 in situ hybridization을 시도하였다. 토끼 출혈증 viral RNA의 보합 결합에 이용하는 probe는 4717에서 4800(84bases)까지 oligonucleotide를 5'말단에 biotin-CE phosphoramidite로 표지하여 사용하였다. 보합결합물의 증명은 신호 표지로서 antibiotin antibody-10mm gold를 사용하였으며, hubridization이나 증명은 기존 protocol에서 약간의 변법을 사용하였다. 0.02% glutaraldehyde에서 고정하고 unicryl resin 포매한 표본, biotinylated oligonucleotide probe, antibiotin antibody-10mm gold로 실험한 결과 증강된 신호를 얻을 수 있었다. 특히 전처리를 생략하므로써 실험 과정을 간단하게 하여 신속한 결과를 얻을 수가 있었다. 전자현미경 in situ hybridization을 통하여 토끼 출혈증 바이러스의 주요 표적은 간세포로 간염 세포의 세포질 내 미토콘드리아와 핵사이에서 immuno gold입자가 뚜렷하게 표지됨으로서 viral RNA를 증명할 수 있었다. Simple stain and electron microscopic in situ hybridization is studied and applied for the identification of rabbit haemorrhagic disease viral RNA in a unicrylated preparation of the liver after innoculation of rabbit haemorrhagic disease virus. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probe labelled by biotin CE-phosphoramidite compared with 4717∼4800 sequences of rabbit haemorrhagic disease virus, modified hybridization protocol and antibiotin antibody-l0nm gold as signal marker. The best results were obtained in 0.02% glutaraldehyde, Unicryl resin cell block, biotinylated oligonucleotide probes, antibiotin-l0nm gold. In this report, RHD viral RNA was distributed widely within the mitochondria and nucleus of liver cell by electron microscopic in situ hybridization. In situ hybridization has become a standard method for localizing DNA or RNA sequences in tissue or cell preparation. In situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method.

In situ analysis of HER2 mRNA in gastric carcinoma: comparison with fluorescence in situ hybridization, dual-color silver in situ hybridization, and immunohistochemistry

Kim, M.A.,Jung, J.E.,Lee, H.E.,Yang, H.K.,Kim, W.H. W. B. Saunders Co ; Centrum Philadelphia 2013 Human pathology Vol.44 No.4

The importance of anti-HER2 therapy has focused attention on the ability of clinical assays to correctly assign HER2 amplification status. In the present study, we evaluated HER2 mRNA expression using a new mRNA in situ detection technique called RNAscope in 211 cases of formalin-fixed, paraffin-embedded gastric carcinoma. In addition, we compared the results with the conventional methods of immunohistochemistry, fluorescence in situ hybridization, and dual-color silver in situ hybridization. RNA in situ hybridization (in situ hybridization) showed that 162 cases (76.8%) were score 0, 5 cases (2.4%) were score 1, 10 cases (4.7%) were score 2, 13 cases (6.2%) were score 3, and 21 cases (10.0%) were score 4. HER2 transcription levels were found to be significantly related to pT class, pN class, and tumor recurrence. mRNA expression was well correlated with protein overexpression and gene amplification; 20 cases out of 23 with DNA amplification showed a score of 4 in RNA in situ hybridization (P < .001). Three cases showed false negative and one case showed false positive results by in situ hybridization. More studies are needed to determine whether the in situ hybridization method can identify additional patients that may benefit from anti-HER2 therapy or exclude those who may be resistant to anti-HER2 therapy.

Klebsiella가 보유한 아미노 배당체 항생제 변경효소 유전자의 분포

이유철,정지인,김정민,이제철,이상화,설성용,조동택 慶北大學校 醫科大學 1994 慶北醫大誌 Vol.35 No.1

목적 :임상 검체에서 분리한 Klebsiella pneumoniae를 대상으로 kanamycin(Km)과 gentamicin(Gm)에 대한 내성양상과 아미노 배당체 항생제 변경효소인 ANT(2") 및 APH(3') 유전자의 분포를 조사하였으며, spot hybridization(SPH)과 plasmid 및 전체 DNA를 Southern hybridization을 시행한 성적을 비교하였다. 재료 및 방법 : 91주의 Klebsiella를 사용하였으며, ANT(2")유전자에 대한 probe는 pFCT3103을 AvaⅠ으로 처리한 후 310bp 크기의 절편을, APH(3')Ⅰ유전자에 대한 probe는 Tn 903을 XhoⅠ과 HindⅢ로 처리한 후 520bp 크기의 절편을 사용하였다. 결과 : 91주 중 20주가 Km과 Gm에 내성을 나타내었으며, 서울에서 분리된 균주의 내성빈도가 29.3%로 대구(11.6%)보다 높았다. 변경효소 ANT(2") prode와 hybridization 양성을 나타낸 것이 13주, APH(3') probe와 hybridization 양성을 나타낸 것이 8주였으며, 2가지 probe 모두에 양성인 것이 4주였는데, APH(3') probe와 hybridization 양성을 나타낸 8주는 모두 서울에서 분리된 균주들이었다. 결론 : 항생제 변경효소의 유전자 probe를 이용하여 hybridization함으로써 단순한 plasmid 크기의 비교나 제한효소 절단 양상의 관찰로 확인하기 힘든 유전자 내용에 대한 조사를 하여 지역간의 항생제 내성의 차이를 증명할 수 있었다. Ninty-one strains of Klebsiella pneumoniae isolated from clinical specimens in Seoul, Taegu, and Pusan areas were evaluated for the resistance to kanamycin(Km) and gentamicin(Gm). Frequency of resistance to aminoglycoside antibiotics of strains isolated in Seoul(29.3%) was higher than those in Taegu(11.6%). Of all strains, 20 strains were resistant to Km and Gm, and harbored 1-4 plasmids ranged from 2.8 to 142 megadaltons (MDa). ANT(2") and APH(3') probes were used to detect aminoglycoside-modifying enzymes ANT(2") and APH (3') genes of these resistant strains. By spot-hybridization, 13 strains were hybridized with both probes. All strains hybridized with APH (3') probes were the Klebsiella pneumoniae strains isolated in Seoul, and 3 strains resistant to Km were hybridized with APH (3') probe. The sizes of plasmids hybridized with ANT(2") were variable and with APH(3') were 82MDa and 17MDa. Four strains harboring 82MDa and 17MDa plasmids were isolated from a hospital in Seoul. 82MDa plasmid of Klebsiella pneumoniae YKK103 was hybridized with both probes. Four strains were hybrdized with the ANT (2") or APH(3')probes in spot-hybridization, but not hybridized with plasmids and chromosomal DNA in Southern-hybridization.

디지털 조형의 혼성적 특성연구

김란희(Kim, Ran-Hee),안성모(Ahn, Seongmo) 한국실내디자인학회 2014 한국실내디자인학회논문집 Vol.23 No.4

The objective of this research is to suggest new geometric possibilities in digital architecture by investigating the characteristics of hybridization in digital geometry. The research begins with theoretical background research such as defining hybridization, investigating hybrid thinking, and studying the theory the theory of digital geometry, along with the four conceptual characteristics of hybridization that could be drawn, such as temporality, liquidity, complexity, and connectivity. Based on these characteristics, the generative method of hybrid digital geometric languages such as Blob, Particle, Morph, Loft, and Boolean was analyzed with case research in contemporary digital architecture. As a result, diverse hybrid geometric keywords were extracted; these keywords suggest potential meanings of hybridization such as accidentality, mobility, diversity, and identity. Different elements represent the “mobility” in time by the force and wave, and they are “accidentally” combined in gradual change. The united species in “diverse” characters are seamlessly connected and emerge as a new “identity.” The research maximizes the generative possibilities in digital geometry and provides a theoretical basis to apply the digital hybrid methods to architectural design by suggesting the potential meanings and possibilities in hybridization.

Hybridization에 의한 반수체 재조합 효모균주의 전분 발효능 증진

박선영,김근,이창후 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6

Hybridization을 통하여 α-amylase와 glucoamylase를 동시에 분비하는 재조합 단수체 효모균주 Saccharomyces diastaticus K114의 전분분해력, 에탄올내성, 당내성, 고온내성등의 발효특성을 증진시키고자 하였다. 이 단수체 효모균주와 glucoamylase의 활성이 좋고 여러가지 발효능이 우수한 단수체 효모균주 S. diastaticus 1177과의 hybridization 결과 얻어진 hybrid HH64주는 에탄올내성, 고온과 당내성이 증진되었으며, 특히 4%의 전분으로부터 1.6%(w/v)의 에탄올을 생산하여, 1.30%(w/v)의 에탄올을 생산한 재조합균주 S. diastaticus K114보다 전분으로부터 에탄올 생성능이 크게 증진되었다. 한편 이 HH64균주의 전분발효에 있어서의 최적온도 및 pH는 각각 30℃와 5이어다. 개발된 hybrid 효모 HH64sms 20%의 전분으로부터 7.5(w/v)의 에탄올을 직접 생산하였다. To improve the fermentation characteristics(such as starch-degradability, ethanol tolerance, sugar and high-temperature tolerance) of recombinant haploid yeast Saccharomyces diastaticus K114, hybridization technique was used. The hybridization partner was S. diastaticus 1177 which had good glucoamylase activity and fermentability. The best hybrid HH64 showed improved ethanol tolerance, sugar and high-temperature tolerance. Especially, the starch-fermentability was significantly improved, since the hybrid produced 1.60% (w/v) ethanol from 4% (w/v) starch, while the recombinant haploid K114 produced 1.30% (w/v) ethanol. The optimum temperature and pH for the starch-fermentation by the hybrid HH64 was 30℃ and 5, respectively. The hybrid yeast HH64 produced 7.5% (w/v) ethanol directly from 20% (w/v) starch.

Melanization plasticity of Drosophila kikkawai, Drosophila leontia and reciprocal hybrids under different temperatures

Singh Divya,Ramniwas Seema,Tyagi Pankaj Kumar,Kumar Girish,Gola Deepak 한국응용곤충학회 2022 Journal of Asia-Pacific Entomology Vol.25 No.1

Drosophila (Sophophora) kikkawai, Burla, 1954 and Drosophila (Sophophora) leontia, Tsacas & David 1978 are closely related sibling species, the former being cosmopolitan and the latter is restricted to tropical localities. We investigated the influence of introgressive hybridization on phenotypic diversity of the two sibling species in the present study. How hybridization supports the relative abundance of pure species according to latitudinal cline is the aim of this study because hybrids show a tendency to acquire geographical location of their parent species in equal or greater abundance. How hybridization supports the plasticity for melanization of hybrids is not explored yet. The two species can cross and generate hybrids. For this, we crossed true breeding strains of both species to obtain the hybrids i.e. dark female (♀) of D. kikkawai (D. k) with males (♂) of D. leontia (D. l) in cross I and light ♀ of D. k with ♂ of D. l in cross II along with their reciprocal crosses. Finally, we studied the plasticity of both species and their hybrids at 6 growth temperatures (14, 17, 21, 25, 28 and 31 ◦ C). We found that there is no plasticity for melanization in true breeding darker and lighter strain of D. kikkawai as well as D. leontia whereas hybrids of both species showed high phenotypic plasticity. Significant differences in slope values across tem peratures in parental and hybrid lines suggest plastic effects. Phenotypic variation in abdominal melanization in hybrids can be interpreted as a result of gene introgression with D. kikkawai. We conclude that introgressive hybridization might be an important, although underestimated, mechanism shaping species distribution and adaptation.

Methicillin 내성 Staphylococcus aureus의 검출을 위한 분자유전학적 기법에 관한 연구

조태흠,김민정,오양효 한국생명과학회 1999 생명과학회지 Vol.9 No.4

Thirty strains of methicillin resistant Staphylococcus aureus were obtained from the clinical isolates. In order to investigate the pursuit of the pathogens of nosocomial infection, these strains were studied for antibiotic sensitivity as well as its resistant pattern. Among the methods of hybridization which directly confirm the specific antibiotic resistant genes by means of the recently developed specific probe DNA, dot blot hybridization and southern blot hybridization were performed and these two methods were compared in their sensitivity and specificity. Strains that is sensitive to cephalothin to the subject of methicillin resistant Staphylococcus aureus were in 43%. Those that are sensitive to cefoperazone and cefuroxime were 26% and 23%, respectively. In case of MIC, MIC50 of cefoperazone was 8 $\mu\textrm{g}$/$m\ell$, and MIC90 was 128 $\mu\textrm{g}$/$m\ell$ to be the lowest. As the results of plasmid DNA electrophoresis, most of methicillin resistant Staphylococcus aureus strains had more than 4 plasmids. These plasmids digested by BamHI, methicillin resistant Staphylococcus aureus is distributed as 10 fragments with the size of 65 kb to 1.5 kb. Dot blot hybridization were performed to examine the existence of mecA gene to show the detection rate of 50%. Southern blot hybridization were done to see if DNA bands which amplify the activity of digoxigenium-labeled probe by PCR were actually PCR products of mecA gene and it showed the detection rate of 53%. It can be concluded that the southern blot hybridization seemed to be better in sensitivity and specificity when it is compared with the results of dot blot hybridization.