• 식물조직배양학회지
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• 제21권2호
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• pp.111-115
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• 1994

벼(Oryza sativa L. cv Nipponbaie)배양세포에 중력 450,000 x g를 처리하여 cDNA library를 만들고, 무처리 및 고중력을 처리한 cDNA 프로브로 스크리닝을 실시하여 고중력에 특이적으로 양성반응을 나타내는 GSC 13 및 GSC124 cDNA를 선발하였다. 선발된 두 유전자 GSC 13 및 GSC 124의 길이는 각각 1.34 및 0.67 kilobase pairs였으며, 배양세포내에서 관련된 transcript의 크기는 각각 2.0 및 1.9 kilobase pairs인 것으로 나타났다. 두 유전자를 프로브로한 Northern hybridization을 실시한 결과 GSC 13, GSC 124 공히 배양세포내에 고중력처리에 의해 특이적으로 축적되는 mRNA가 나타났으며, 중력강도 300,000 x g 에 비해 450,000 x g 처리에서 더욱 강한 축적을 보였고 450,000 x g 4시간 처리에서 최대의 수준을 보였다.

• 한국균학회지
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• 제27권3호통권90호
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• pp.187-190
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• 1999

팽이버섯의 자실체 분화 과정에서 특이적으로 발현하는 유전자 분리를 위한 cDNA library는 발이처리 후 7일째 배양한 균사체의 mRNA에 의해 만들어졌다. cDNA클론 FVFD16(Flammulina velutipes fruiting body differentiation)은 자실체 분화 과정에서 특이적으로 발현되는 클론으로 differential screening에 의해 선발되었다. Northern 분석에 의해 FVFD16의 발현 특성을 관찰한 결과, 1일과 4일째의 균사체에서 현저한 발현량을 나타내었다. FVFD16의 염기 서열을 검색한 결과, FVFD16의 mRNA는 open reading frame을 포함한 128의 아미노산 잔기(13.5kDa)를 가진 단백질로 추정되었다.

• Molecules and Cells
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• 제27권5호
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• pp.577-582
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• 2009

The development of a high-throughput functional genomic screening provides a novel and expeditious approach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regulates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ${\gamma}$, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.

• Journal of Life Science
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• 제9권1호
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• pp.14-16
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• 1999

Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2063-2068
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• 2012

Aim: Liquid-based cytology is the most often used method for cervical cancer screening, but it is relatively insensitive and frequently gives equivocal results. Used as a complementary procedure, the high-risk human papillomavirus (HPV) DNA test is highly sensitive but not very specific. The human telomerase RNA gene (TERC) is the most often amplified oncogene that is observed in cervical precancerous lesions. We assessed genomic amplification of TERC in liquid-based cytological specimens to explore the optimal strategy of using this for cervical cancer screening. Methods: Six hundred and seventy-one residual cytological specimens were obtained from outpatients aged 25 to 64 years. The specimens were evaluated by the Digene Hybrid Capture 2 (HC2) HPV DNA test and fluorescence in situ hybridization (FISH) with a chromosome probe to TERC (3q26). Colposcopic examination and histological evaluation were performed where indicated. Results: The TERC positive rate was higher in the CIN2+ (CIN2, CIN3 and SCC) group than in the normal and CIN 1 groups (90.0% vs. 10.4%, p < 0.01). In comparison with the HC2 HPV DNA test, the TERC amplification test had lower sensitivity but higher specificity (90.0% vs. 100.0%, 89.6% vs. 44.0%, respectively). TERC amplification test used in conjunction with the HC2 HPV DNA test showed a combination of 90.0% sensitivity and 92.2% specificity. Conclusion: The TERC amplification test can be used to diagnose cervical precancerous lesions. TERC and HPV DNA co-testing shows an optimal combination of sensitivity and specificity for cervical cancer screening.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.396-397
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• 2005

The polyene antibiotics are a family of most promising antifungal polyketide compounds, typically produced by actinomycetes species. Using the polyene CYP-specific PCR screening with served actinomycetes genomic DNAs, Pseudonocardia autotrophica strain was identified to contain a unique polyene-specific CYP gene. The genomic DNA library screening using the polyene-specific CYP gene probe revealed the positive cosmid clone containing an approximately 34.5 kb DNA fragment revealed a total of seven complete and two incomplete open reading frame (ORFs), which are highly homologous but unique to previously-known polyene biosynthetic genes. These results suggest that the polyene-specific screening approach should be an efficient way of isolating potectially-valuable cryptic polyene biosynthetic gene cluster from various rare actinomycetes.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1022-1027
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• 2005

Based on plasmid display technology by the complexes of fusion protein and the encoding plasmid DNA, an in vitro selection method for high affinity DNA-binding protein was developed and experimentally demonstrated. The GAL4 DNA-binding domain (GAL4 DBD) was selected as a model DNA-binding protein, and enhanced green fluorescent protein (EGFP) was used as an expression reporter for the selection of target proteins. Error prone PCR was conducted to construct a mutant library of the model. Based on the affinity decrease with increased salt concentration, mutants of GAL4 DBD having high affinity were selected from the mutant protein library of protein-encoding plasmid complex by this method. Two mutants of (Lys33Glu, Arg123Lys, Ile127Lys) and (Ser47Pro, Ser85Pro) having high affinity were obtained from the first generation mutants. This method can be used for rapid in vitro selection of high affinity DNA-binding proteins, and has high potential for the screening of high affinity DNA-binding proteins in a sequence-specific manner.

• 약학회지
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• 제42권2호
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• pp.144-148
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• 1998

One hundred and seventeen crude drugs were screened for DNA-binding activity in vitro. The DNA-methyl green assay is a useful biochemical screen for the detection or development of biologically active compounds which bind to DNA. This assay is based upon the fact that free methyl green undergoes rapid spontaneous molecular rearrangement to its colorless carbinol base so that the liberation of the dye from DNA by displacement can be followed spectrophotometrically as a decrease in absorbance at 65Onm. Seven methanolic extracts of crude drugs including Cinnamomi Cortex spissus, Coicis Semen, Coptidis Rhizoma, Perillae Semen, Plantaginis Semen, Polygalae Radix and Zanthoxyli Fructus showed less than 2,000${\mu}$g/ml as their $IC_{50}$ values. The active principles of Plantaginis Semen and Zanthoxyli Fructus were transferred into organic solvents, which showed the $IC_{50}$ values with 588 and 574${\mu}$g/ml, respectively. These fractions have been selected for isolation of biologically active constituents.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8495-8501
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• 2016

Purpose: This study was conducted to assess the agreement and differences between cervical self-sampling with a Kato device (KSSD) and gynecologist sampling for Pap cytology and human papillomavirus DNA (HPV DNA) detection. Materials and Methods: Women underwent self-sampling followed by gynecologist sampling during screening at two primary health clinics. Pap cytology of cervical specimens was evaluated for specimen adequacy, presence of endocervical cells or transformation zone cells and cytological interpretation for cells abnormalities. Cervical specimens were also extracted and tested for HPV DNA detection. Positive HPV smears underwent gene sequencing and HPV genotyping by referring to the online NCBI gene bank. Results were compared between samplings by Kappa agreement and McNemar test. Results: For Pap specimen adequacy, KSSD showed 100% agreement with gynecologist sampling but had only 32.3% agreement for presence of endocervical cells. Both sampling showed 100% agreement with only 1 case detected HSIL favouring CIN2 for cytology result. HPV DNA detection showed 86.2%agreement (K=0.64, 95% CI 0.524-0.756, p=0.001) between samplings. KSSD and gynaecologist sampling identified high risk HPV in 17.3% and 23.9% respectively (p=0.014). Conclusion: The self-sampling using Kato device can serve as a tool in Pap cytology and HPV DNA detection in low resource settings in Malaysia. Self-sampling devices such as KSSD can be used as an alternative technique to gynaecologist sampling for cervical cancer screening among rural populations in Malaysia.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.315.1-315.1
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• 2002

DNA topoisomerases I (topo I) and II are essential enzymes that relax DNA supercoiling and relieve torsional strain during DNA processing. including replication. transcription. and repair. Topo I relaxes DNA by cleaving one strand of DNA by attacking a backbone phosphale with a catalytic lyrosine (Tyr723. human topo I). This enzyme has recently been investigated as a new target for antineoplastic drugs. Inhibitors to the enzyme intercalate between the DNA base pairs. interfering religation of cleaved DNA, therefore inhibit the activity of topo I. (omitted)