• Biomedical Science Letters
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• v.10 no.2
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• pp.75-84
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• 2004

Massive identification of differentially expressed patterns has been used as a tool to detect genes that are involved in disease related process. We employed circular single stranded sense molecules as probe DNA for a DNA chip. The circular single stranded DNAs derived from 1,152 unigene cDNA clones were purified in a high throughput mode from the culture supernatant of bacterial transformants containing recombinant phagemids and arrayed onto silanized slide glasses. The DNA chip was examined for its utility in detection of differential expression profile by using cDNA hybridization. Hybridization of the single stranded probe DNA were performed with Cy3- or Cy5-labeled target cDNA preparations at $60^\circ$C. Dot scanning performed with the hybridized slide showed 29 up-regulated and 6 down-regulated genes in a cancerous liver tissue when compared to those of adjacent noncancerous liver tissue. These results indicate that the circular single stranded sense molecules can be employed as probe DNA of arrays in order to obtain a precious panel of differentially expressed genes.

• Korean Journal Plant Pathology
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• v.10 no.1
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• pp.1-6
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• 1994

Randomly chosen recombinant clones of Fusarium oxysporum were analysed to select useful probes for relatedness analysis of Fusarium oxysporum. Genomic DNA of F. oxysproum f. sp. cubense, digested with HindIII, was ligated to pUC118 and used to transform Escherichia coli strai DH5$\alpha$. Three clones were identified that hybridized to mutiple restriction fragments of some formae speciales of F. oxysporum. These probes detected repetitive sequences in HindIII or EcoRI digested DNAs. Repeated copy clone pFC46, pFC52 and pFC54 showed evident polymorphisms among ten formae speciales of this fungus. Since clone pFC 52 strongly hybridized to multiple EcoRI-digested restriction fragments of f. sp. cubense, it may be useful as a probe for analysis of other genetic characteristics of this forma specialis. The results suggest that our clones might be very useful as probes for relatedness analysis between or within formae speciales of Fusarium oxysporum.

• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.362-368
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• 1997

Five formae speciales of Fusarium oxysporum in Korea were examined using RFLP analysis to find the possibility for classification and analyze genetic variations. DNAs from F. oxysporum f. sp. lycopersici, cucumerinum, fragariae, garlic and sesami were used with three recombinant probes such as pFC46, pFC52 and pFC57. Distinct differences among five formae speciales of this fungus were detected in RFLP band patterns based on southern hybridization of genomic DNA using each recombinant clone, which was a repetitive copy probe. Strains belong to four formae speciales could be very stable in genetic variation except f. sp. sesami which has more variation than the others based on the RFLP analysis. They formed their own cluster which has high similarity within the same formae specialis resulted from the UPGMA analysis for genetic relationship analysis and each cluster represented its own formae specialis. The method using three recombinant DNA probes could be a good tool for classification of formae speciales in F. oxysporum.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.116-121
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• 1987

In order to cloning of the nif genes of Enterobacter agglomerans NFB-264, the digested total DNA of the strain was ligated to pBR 322 and transformed into E. coli. Through the negative selection and colony hybridization, the transformants were obtained. The recombinant plasmids, pNEL 10 and pNES 20 were extracted from these transformants. It was known from Southern hybridization that pNEL 10 contained the 12 Mdal foreign DNA fragment hybridized with nif Q-X probe and pNES 20 included the 5 Mdal foreign DNA fragment hybridized with nif NE and nif YK probe.

• Microbiology and Biotechnology Letters
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• v.14 no.2
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• pp.133-137
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• 1986

17S-ribosomal RNA gene from the hygromycin resistant protozoan Tetrahymena thermophila hmr 3 was cloned on E. coli vector pBR 322 as part of study to work the 17S-rRNA structure and the mechanism of hygromycin resistance. The 17S-rDNA was inserted into the Hind 111 site of pBR 322. The clones having recombinant plasmid were selected by the method of colony hybridization with a 17S-rDNA probe of wild type B1868. The orientation of 17S-rDNA insert was located near the tetracycline resistant gene of pBR 322 in a clone 5-19 with the recombinant plasmid.

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.389-398
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• 1992

Molecular cloning was carried out on the Danish strain of bovine viral diarrhea virus(BVDV) to construct strategy for the diagnostic tools and effective vaccine of BVD afterwards. A recombinant DNA clone(No. 29) was established successfully from cDNA for viral RNA tailed with adenine homopolymer at 3'-end. $^{32}P$-labeled DNA probes of 300~1,800bp fragments, originating from the clone 29, directed specific DNA-RNA hybridization results with BVDV RNA. Recombinant DNA of the clone 29 was about 5,200bp representing 41.6% of the full length of Danish strain's RNA, and restriction sites were recognized for EcoR I, Sst I, Hin d III and Pst I restriction enzymes in the DNA fragment.

• BMB Reports
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• v.28 no.3
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• pp.216-220
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• 1995

cDNA for human cholesteryl ester transfer protein (CETP), a potent atherogenic plasma protein that redistributes the neutral lipids among lipoproteins, was expressed in recombinant vaccinia virus-infected cells (CV-1). Two insertion vectors regulated by different promoters were constructed. The vectors were introduced into human thymidine kinase-negative ($TK^-$) 1438 cells infected with wild-type vaccinia virus (WR strain). Recombinant viruses were selected with 5-bromodeoxyuridine (BUdR) and X-gal and identified with DNA dot blot analysis (vSC11-CETP and vTM1-CETP). The CETP cDNA insert in the recombinant vaccinia virus genome was identified by Southern blot analysis. Transcription of CETP cDNA in CV-1 cells infected with recombinant vaccinia virus was monitored by Northern blot analysis using the CETP cDNA as a probe. Positive signals were detected at 1.8 kb in cells infected with vSC11-CETP and at 2.3 kb in cells infected with vTM1-CETP. The recombinant vaccinia virus-infected CV-1 cells were shown to produce functional CETP when the culture medium was subjected to the CETP assay.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.281-290
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• 2002

The purpose of this study is to develop species-specific DNA probe for detection and identification of Prevotella intermedia (P. intermedia) G8-9K-3. This study procedure includes (1) whole-genomic DNA extraction of P. intermedia G8-9K-3 (2) construction of the genomic DNA library, (3) screening of strain-specific DNA probe by reverse dot hybridization, (4) confirmation of strain-specific DNA probe by Southern blot hybridization, (5) determination of nucleotide sequences of strain-specific DNA probe. Twenty-eight recombinant plasmids containing Hind III-digested DNA fragments of P. intermedia G8-9K-3 were obtained. Reverse Dot Hybridization and Southern blot analysis data showed that one of them, Pig3, could be P. intermedia G8-9K-3-specific DNA probe. This datum indicates that this Pig3 DNA probe could be useful in detection and identification of the P. intermedia G8-9K-3 strain.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.3
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• pp.275-282
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• 1993

The objective of this study was to clone a partial fragment of $\alpha$-amylase from Korean maize. We designed and synthesized an oligonucleotide probe and two kinds of PCR primers based on cDNA conserved region of $\alpha$-amylase sequences from other plants. Total RNA from 3-day-old maize seedling was used as template for 1st strand cDNA synthesis and RNA-DNA hybrid was used as template for polymerase chain reaction(PCR). The product of PCR was about 0.5 kb long and inserted into pUC19. We named this recombinant plasmid as pZM$\alpha$'. The cloned fragment was certified by Southern blot analysis using labeled synthetic oligonucleotide as probe.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.248-256
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• 1996

The present study was performed to clone and express human poly (ADP-ribose) synthetase (PARS) cDNA in E coli. For these purposes, the CDNA for human poly (ADP-ribose) synthetase, encoding the entire protein, was cloned into pGEM-7Zf(+). The resulting recombinant plasmid pPARS6.1 was restriction enzyme mapped and its identity was confirmed by Southern blot analysis. The pPARS6. 1 contained full-length CDNA of human PARS and the nudeotide sequences were identical with those reported previously. The recombinant protein which migrated as a unique 120 kDa band on 10% SDS-polyacrylamide gels, was identified as PARS by Southwestern blots using nick-translated DNA probes and by activity gels and activity blots using 32 P-NAD as a substrate for poly (ADP-ribose) synthetase (PARS). The signals corresponding to 120 and 98 kDa proteins were obtained following IPTG (0.4 mM) induction of the PARS cDNA cloned into Xba I-digested pGEM-7Zf(+) vector. Nonspecific signals corresponding to 45 and 38 kDa proteins were also shown in both IPTG-induced and noninduced cells. The nonspecific proteins may be products of incomplete translation or proteolytic products of intact PARS.