• Title/Summary/Keyword: cDNA

Search Result 6,234, Processing Time 0.031 seconds

Effects of Ara-C on UV and MMS-induced Excision Repair, Chromosome Aberrations, Sister Chromatid Exchanges and Replication Inhibition (자외선과 MMS에 의한 절제회복, 염색체이상, 자매염색분체 교환 및 복제억제 현상에 미치는 Ara-C의 영향)

  • Park, Kyung-Hee;Park, Sang-Dai
    • The Korean Journal of Zoology
    • /
    • v.23 no.4
    • /
    • pp.203-218
    • /
    • 1980
  • Unscheduled DNA synthesis, chromosome aberrations, sister chromatid exchanges and DNA replication inhibition induced by the combined treatments with ara-C and UV-light or MMS in $HF_1$, CHO and $HelaS_3$ cells were studied, and the results obtained were as follows: (1) Ara-C was found to inhibit UV-or MMS-induced unscheduled DNA synthesis and the inhibitory effect of ara-C was more remarkable in its post-treatment. (2) Ara-C enhanced the rate of chromosome aberrations induced by MMS or UV-light. Post-treatment with ara-C exhibited the synergistic effect on MMS-induced chromosome aberrations mainly by increases of chromatid deletions. (3) Contrarily, ara-C did not increase the rate of sister chromatid exchanges, particularly in the pre-treatment with MMS, although it was found to induce sister chromatid exchanges. (4) The rate of DNA synthesis was declined immediately after are-C treatment and then recovered. The combined treatments with ara-C and UV-light or MMS showed that the initial response on replication inhibition was similar to that of ara-C, but later responses were similar to that of UV-light or MMS treated group.

  • PDF

Cloning and Expression of Human Clotting Factor 9 cDNA un Escherichia coli (인체 혈액응고 9인자 cDNA cloning 및 Escherichia coli 에서의 발현)

  • Young Won Lee;Hyang Suk Hur;Myoung Hee Kim
    • Biomedical Science Letters
    • /
    • v.2 no.2
    • /
    • pp.231-240
    • /
    • 1996
  • Human blood clotting (coagulation) factor 9 cDNA which codes for 461 amino acid has been cloned by screening human fetal liver cDNA library using PCR. This 1.4 kb cDNA spanning from the ATG initiation codon to the TAA termination codon was cloned into bacterial .expression vector pGEX-2T, generating pGEX-F9 plasmid. The plasmid pGEX-F9 expresses about 73 kDa GST (Glutathione S-transferase)-Factor 9 fusion protein when introduced into E. coli. Western blot analysis using polyclonal antibody raised against human factor 9 confirmed this fusion protein contains factor 9 protein. The level of GST-factor 9 expression was about 20% of total protein and the purification of fusion protein was efficiently achieved by using GST agarose bead based on one step purification protocol.

  • PDF

Campylobacter jejuni 의 열충격 반응과 그유전자에 관한 연구

  • 김치경;임채일;이길재
    • Korean Journal of Microbiology
    • /
    • v.30 no.3
    • /
    • pp.232-238
    • /
    • 1992
  • Canz~~j~lohuc;tc.~jurn i werc studied for their heat shock responses at several elevated temperatures and their heat shock genes were detected by the technique of Southern hybridization. (.. ,jc\ulcorneruni sy~>thesized the major heat shock proteins of hsp90. hsphh. and hsphO at 48$^{\circ}$C . ant1 their w~u.ival rates were maintained as the same level at optimal temperature. '1-hc heat shock genes in chromosome of C ,jc:jutii werc determined to be homologous to the heat shock genes or E. t,oli. by showing strong signals in Southern hybridization analysis using clnaK and groESL- as DNA probe But the restriction sites for thc fragmcnts including heat shock genes were different betueen E. c,oli and C ,jtjuni.

  • PDF

restriction Site Polymorphism of mtDNA for differentiating Anopheles quadrimaculatus (Say) Sibling Species (미토콘드리아 DNA 제한효소 절단부위 변이에 의한 Anopheles quadrimaculatus (Say) 모기의 자매종 구별)

  • ;S.K. Narang
    • Korean journal of applied entomology
    • /
    • v.29 no.2
    • /
    • pp.132-135
    • /
    • 1990
  • Three mitochondrial cDNA probes from Aedes albopictus were used to demonstrate restriction site polymorphism in mtDNA of three sibling species of Anopheles quadrimaculatus(Say). It was shown by DNA hybridization to have substantial sequence homology betwen the mtDNA of different genus. The proves reveled local restriction site variation between members of the Anopheles quadrimaculatus sibling species complex. Mitochondrical DNA (mtDNA), isolated from individual mosquitoes was digested by type II restriction enzymes and four enzymes were found to be useful for the purpose. Hind III alone could be used to obtain a diagnostic restriction pattern.

  • PDF

Partial Cloning of Histone Deacetylase Genes from Ganoderma lucidum. (영지에서 Histone Deacetylase 유전자의 부분 클로닝)

  • Kim Sunkyung;Kum Joohee;Choi Hyoung T.
    • Korean Journal of Microbiology
    • /
    • v.40 no.3
    • /
    • pp.226-229
    • /
    • 2004
  • Histone deacetylase (HDAC) removes acetyl group in lysine residue of histone protein, which is transferred by histone acetylase. HDAC is important in the stabilization and regulation of gene expression in eukaryotic organisms. PCR has been carried out to clone HDAC genes using cDNA library and genomic DNA as the templates from Ganoderma lucidum isolated in Korea. One 470 bp cDNA gene fragment, and 3 genomic HDAC fragments (585 bp, 589 bp, 630 bp) were amplified. When their deduced amino acid sequences were compared with other fungal HDACs, they showed 59-72% homology.

Isolation of a cDNA Encoding a Chloroplast Triosephosphate Isomerase from Strawberry

  • Kim, In-Jung;Lee, Byung-Hyun;Jinki Jo;Chung, Won-Il
    • Journal of Plant Biotechnology
    • /
    • v.2 no.3
    • /
    • pp.115-121
    • /
    • 2000
  • A cDNA clone encoding chloroplast triosephosphate isomerase (TPI-cp) was isolated from strawberry fruit cDNA library. Sequence analyses indicated that the cDNA contains an open reading frame of 314 amino acids (33.5 kDa) composed of a transit peptide (59 amino acids) in amino terminal region and mature protein (255 amino acids). The existence of transit peptide in the deduced amino acid sequence implies that it encodes a chloroplast isoform. The protein sequence is more similar to other plant chloroplast isoforms than cytosolic isoforms. RNA blot analysis indicated that its expression is ubiquitous in examined five tissues, flowers, leaves, petioles, roots and fruits, and shows differential pattern according to fruit ripening. Genomic DNA blot analysis showed that TPI-cp is encoded by multiple genes in strawberry. Through sequence comparison and phylogenetic tree construction, TPI-cp is distinctively grouped into dicot and chloroplast isoforms.

  • PDF

Detection of DNA Adduct Formed by Mitomycin C by $^{32}P$-Postlabelling ($^{32}P$-Postlabelling 방법을 이용한 미토마이신 C에 의하여 형성된 DNA adduct의 검출)

  • Jeong, Hye-Yun;Kim, Jae-Hyeon;Park, Chang-Won;Lee, Dong-Gwon
    • YAKHAK HOEJI
    • /
    • v.40 no.4
    • /
    • pp.442-448
    • /
    • 1996
  • Mitomycin C(MMC) has been used as an anticancer drug and behaves as an alkylating agent forming covalent cross-link between complementary strands of double strand DNA. The purpose of this research was to determine number of DNA adducts, formed in vivo by Mitomycin C, in mouse organs. DNAs from liver, lung, brain and pancreas were isolated and used for $^{32}P$-postlabelling. The labeled nucleotides were separated by 2D-TLC and subjected to autoradiography. Numbers of MMC-DNA adducts were 9,9,5,4 in liver, pancreas, lung and brain, respectively.

  • PDF

Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain (토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석)

  • 이청호;이영기;강신웅;박은경
    • Journal of the Korean Society of Tobacco Science
    • /
    • v.18 no.2
    • /
    • pp.101-106
    • /
    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

  • PDF

Identification of Radiation-Sensitive Gene in U937 Cell by using cDNA-Chip Composed of Human Cancer Related Gene (U937 세포에서 발암관련 유전자들로 구성된 DNA chip을 이용한 방사선 감수성 유전자들의 선발)

  • 김종수;김인규;강경선;윤병수
    • Environmental Mutagens and Carcinogens
    • /
    • v.22 no.1
    • /
    • pp.54-59
    • /
    • 2002
  • We have used cDNA microarray hybridization to identify gene regulated in response to gamma-irradiation in U-937 cell. The cDNA-chip was composed entirely of 1,000 human cancer related gene including apoptosis and angiogenesis etc. In gamma-irradiated U-937 cell, highly charged protein, ribosomal protein L32, four and a half LIM domains 3, lipocalin 2 (oncogene 24p3) and interleukin 15, ataxia telangiectasia mutated (includes complementation groups A, C and D) genes showed increased level of its transcription, and cell division cycle 25A, dihydrofolate reductase, topoisomerase (DNA) II beta(180kD), kinase suppressor of ras and strarigin genes showed reduced level of its transcription compared to untreated U-937 cell. The significant change of level of transcription was not found in well-known ionizing radiation(IR)-responsive gene, such as transcription factor TP53 and p53 related gene, except ataxia telangiectasia mutated gene.

  • PDF

Acquisition of Thermotolerance in the Transgenic Plants with BcHSP17.6 cDNA (BcHSP17.6 cDNA의 도입에 의한 형질전환된 식물의 내열성 획득)

  • Ki Yong Kim;Min Sup Chung;Jin Ki Jo
    • Journal of The Korean Society of Grassland and Forage Science
    • /
    • v.17 no.4
    • /
    • pp.379-386
    • /
    • 1997
  • Recombinant plasmid, pBKH4, containing NPT II and P35S-BcHSP17.6 was constructed by ligation of Bum H I -digested pBKSl-l and BcHSP 17.6 (thermotolerance gene) 6om pBLH4. The tobacco leaf disc was cocultivated with transformed Agmbacterium tumefaciens bearing pBKH4 for 24 hours and transformed shoots were selected on MS-n/B medium containing $100\;{\mu\textrm{g}}/ml$ of kanamycin. Heat-killing temperature of Nicotima tabacum was $50^{\circ}$ for >15min, and transformed tobacco plants with BcHSP17.6 cDNA exhibited thermotolerance at the heat-killing temperature. The transgenic plants were analyzed by Southern blot hybridization with the probe of ${\alpha}^{_32}P$ labelled BcHSP17.6 cDNA. Transcription and expression level of BcHSP17.6 cDNA were also continued by Northern blot analysis and Ouchterlony double immunodiffusion assay. In this study, we suggest that the BcHSP17.6 cDNA introduced to tobacco plant is related to thenuoto-lerance and 17.6-kD LMW HSP acts as a protector from heat damage in plants.

  • PDF