• The Korean Journal of Zoology
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• v.33 no.1
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• pp.35-44
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• 1990

Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.

• Animal cells and systems
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• v.1 no.4
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• pp.629-635
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• 1997

Previously we reported the migration and rearrangement of a chloroplast gene cluster into mitochondria. The exact genomic locations of the clusters, modes of the gene rearrangement and mechanisms of the interorganellar migration of the clusters have yet to be understood. The detailed analysis needs to include a larger region of DNA surrounding each cluster. To study DNA rearrangement and migration in more detail a cosmid library was constructed using the total rice genomic DNA including nuclear, chloroplast and mitochondrial DNA. From this cosmid library, a sub-library was obtained by selecting the clones hybridized to various regions of chloroplast DNA. According to the hybridization pattern 136 clones from the sub-library were classified into 29 groups. Detailed analysis of these clones revealed that in addition to authentic chloroplast DNA, the clones contain its homologs resulted from rearrangement and mutation. We analyzed two clones in detail, which contain different rp12 homologs resulted from rearrangement and/or migration, respectively.

• Korean Journal of Cognitive Science
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• v.1 no.2
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• pp.361-376
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• 1989

Norepoine is N-methylated by the enzyme phenly ethanolamine N-metyltransferase(PNMT)to form epinephrine.this enzyme is larhly restructed to the adrenal medulla where epinephrine in mammalian brain where epinephrine function as a neurotransmitter.It seems clear that central epinephrine is involved in the regulation of cardiovacular function and in several forms of hypertension.However,information about the struture of mammalian epinephrine forming enzyme has been limited until now.But recently we isolate bovine and human PNMT cDNA clone using gtll expression library and sequcde total nucleotide composition.To obtain information about the structrue of the human and rat PNMT proteins and gones and to further define the extent of the evolutionary relationships among the PNMT molecules of these species human and rat genomic DNA clones to PNMT were sequentially isolated and characterized.

• Archives of Pharmacal Research
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• v.19 no.6
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• pp.450-455
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• 1996

The genomic DNA was prepared from trout liver which was treated with 3-methycholanthrene, and cloned into lambda EMBL3 at BamHl site. The genomic library was constructed via infections of these recombinant phages into E. coli K802, and screened by the most $5^I$-portion of trout CYP1A1 cDNA. After the screening of $10^9$ clones of the amplified library, 12 positive clones were isolated, and subjected to further screenings. The results of southern blot hybridization of genomic DNA prepared from the positive clone showed the presence of a single gene of CYP1A1, and 3.5 Kb PstI fragment that hybridizes with the most $5^I$-region DNA of CYP1A1 cDNA. The restriction map of PstI fragment was determined by the restriction digestion with various enzymes. The nucleotide sequence of the upstream genomic DNA of CYPIAI was determined by DNA sequencing of exonuclease III unidirectionally deleted PstI fragment DNA using $[^{35}/S]$dATP. This paper presented the upstream genomic DNA of CYP1A1 contained a part of coding region which was about 351 base pairs (from ATG to PstI site at 3563).

• Korean journal of applied entomology
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• v.36 no.1
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• pp.96-104
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• 1997

A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

• The Korean Journal of Mycology
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• v.30 no.1
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• pp.66-72
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• 2002

To develop DNA markers for analysis of genetic characteristics of Phytophthora capsici population, randomly selected clones from HindIII-digested genomic DNA library of P. capsici 95CY3119 were surveyed by hybridizing to Southern blots of HindIII-digested total genomic DNA of P. capsici. Probe DNAs inserted into selected individual clones strongly hybridized with HindIII digests of P. capsici. Among probes examined, PC9 revealed the repetitive and highly polymorphic bands to HindIII digests of inter-and intra-field P. capsici isolates. Genetic diversity of individual isolates was also clearly revealed in cluster analysis based on its band patterns. The other probe, PC22, was hybridized only to DNA from P. capsici and this was highly repetitive. However, there was no response to other Phytophthora species and Pythium sp. These DNA probes could be used as very useful markers in analysing genetic diversity and identification for P. capsici population throughout the world.

• Mycobiology
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• v.38 no.1
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• pp.70-73
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• 2010

Temperature-sensitive yeast mutants were used to screen for cell cycle-related genes from Pleurotus eryngii genomic DNA. A mushroom genomic DNA library was established and each gene was screened for the ability to rescue seven Saccharomyces cerevisiae temperature-sensitive strains. Hundreds of yeast transformants were selected at restrictive temperatures over $30^{\circ}C$. Plasmids from the transformants that survived were isolated and transformed back into their host strains. The temperature sensitivity of the resulting transformants was tested from $30^{\circ}C$ to $37^{\circ}C$. Ten DNA fragments from P. eryngii were able to rescue yeast temperature-sensitive strains, and their DNA sequences were determined.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.660-664
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• 2000

To clone genes related UK-58,852 production, genomic DNA of strain Actinodura roseorufa was used for the construction of genomic library using pOJ446 cosmid vector. The genomic library was screened rising dehydratase PCR product and eryA gene as a DNA hybridization probe. pHD54 was isolated, which contained an approximately 35kb of inserted DNA. BamHI, SmaI and sonicater fragments hybridized to eryA probe. All of pHD54 BgmHI, SmaI and sonicater fragments were subcloned into pGEM7 and some fragments which hybridized to eryA probe were sequenced. The nucleotide sequence was analysed using BLAST program. The sequence identities were observed in KS,AT, KR, ER and PKS loading domains. Also oxidoreductase showed similarity to rifamycin module10, and dTDP-D-glucose 4,6 dehydratase and TDP-D-glucose synthase involved in biosynthesis of sugar showed similarity to Streptomyces argillaceus.

• Korean Journal of Veterinary Research
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• v.34 no.2
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• pp.387-394
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• 1994

To make the genomic DNA probe of Theileria sergenti, the merozoites were purified from bovine erythrocytes. The infected erythrocytes were lysed by Aeromonas hydrophila(Ah-1) hemolysin, and the parasites were isolated by ultracentrifugation on a Percoll discontinuous density gradient. For construction of a T sergenti genomic DNA library, T sergenti DNA was digested with Pstl and the fragments were ligated into the PstI site of pUC19 before transformation of Escherichia coli JM83. Out of thousands of transformants obtained by transformation of E coli JM83 with the genomic library, three plasmids were chosen. The sizes of the inserted DNAs were 2.9kb(2.4kb and 0.5kb) in pKTS1, 4.3kb in pKTS2 and 1.5kb in pKTS3, respectively. The DNA fragments used as probe KTS1(2.4kb), KTS2(4.3kb) and KTS3(1.5kb) were labeled digoxigenin-11-dUTP for the Southern hybridization. In Southern hybridization, all of the probes(KTS1, KTS2 and KTS3) reacted specifically to T sergenti DNA, but not to bovine leucocyte DNA. In order to find out the sensitivities of the digoxigenin-11-dUTP-labeled KTS1 and KTS3 as the probes, purified merozoite DNA and bovine DNA (control) were checked by dot blot hybridization with the probes. Both of the probes, KTS1 and KTS3, detected as minimum amount of 975pg of the T sergenti DNA, but not bovine DNA even to 500ng.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.228-234
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• 1994

Genomic RNA was extracted from Cy strain of odontoglossum ringspot tobamovirus (ORSV-Cy) isolated from infected leaves of tobacco cv. Samsun. Size of the genomic RNA was about 6.6 kb in length. The genomic RNA was fractionated using Sephadex G-50 column chromatography into 2 fractions. They were polyadenylated at their 3'-end using E. coli poly(A) polymerase. Polyadenylated viral RNA was recovered by oligo (dT) primer adapter containing NotI restriction site and Moloney murine leukemia virus SuperScript reverse transcriptase (RNase H-). Second-strand cDNA was synthesized by using E. coli DNA ligase, E. coli DNA polymerase I and E. coli RNase H. Recombinant plasmids containing cDNAs for ORSV-Cy RNA ranged from about 800 bp to 3,000 bp. Among the selected 238 recombinants, pORCY-124 clone was the largest one covering 3'-terminal half of the viral RNA. This clone contained two restriction sites for EcoRI and XbaI and one site for AccI, AvaI, BglII, BstXI, HindIII, PstI, and TthIII 1. respectively. The clone contained partial viral replicase, a full-length movement protein and a complete coat protein genes followed by a 3' untranslated region of 414 nucleotides based on restriction mapping and nucleotide sequencing analyses. Clones pORCY-028, -068, -072, -187 and -224 were overlapped with the pORCY-124. Clones pORCY-014 and -095 covered 5' half upstream from the middle region of the viral RNA, which was estimated based on restriction mapping and partial sequence analysis. Constructed cDNA library covered more than 90% of the viral genome.