• Journal of Microbiology and Biotechnology
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• 제5권1호
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• pp.1-5
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• 1995

A genomic DNA of alkaliphilic bacterium, Micrococcus sp. Y-l, was analysed using the physical mapping method of pulsed-field gel electrophoresis (PFGE). Five restriction enzymes of Sspl, Hpal, Xbal, Ndel or EcoRI, which recognize the Adenine-Thymine-rich sequences of genomic DNA, were used for the generation of few (7 to 20) distinctly separate fragments, with average sizes in the range of 200～500 kb. However, the sites for Notl and SfiI, 8 base-recognizing enzymes, were highly frequent. The genome size of this strain was determined to be 4 mega base pairs (Mb) from restriction fragments separated by PFGE. This is the first case of restriction mapping in alkaliphilic bacterium.

• 한국식물병리학회지
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• 제10권3호
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• pp.235-239
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• 1994

Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

• 한국동물위생학회지
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• 제27권1호
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• pp.103-109
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• 2004

Babesia equi ema-1 5' intergenic(IG) nucleotide was PCR amplified and analyzed for restriction sites in order to identify a promoter region in this IG nucleotide sequence. B equi ema-1 5' IG specific primers identified a 1268 bp PCR product. The sequence had restriction sites for 34 restriction enzymes when analyzed by a computer program. Among them, 26 enzymes had only one restriction site, but the others had more than one sites. When four restriction enzymes (Bgll , HindⅢ, Kpn1 and BamH1) were treated to digest the 1268 bp nucleotide, they had restriction sites as expected by the computer program. Information of restriction sites in the 1268 bp IG nucleotide will be applied to select restriction enzymes for cloning the IG nucleotide to a vector.

• 한국식물병리학회지
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• 제10권3호
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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.

• 약학회지
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• 제36권3호
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• pp.255-258
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• 1992

The clinical isolate Staphylococcus aureus SA8 was resistant to tetracycline(Tc) and harboured a plasmid pKH1(24.82 kb). pKH1 was shown by curing and by transformation to specify resistance to Tc. The cleavage map of a pKH1 was determined by restricction enzyme mapping techniques. Cleavage map is given for BglII, EcoRI, HpaII, PvuII and SalI. Restriction endonuclease BamHI, BglI, BstEII, HpaI, PstI, and XhoI have no sites on this plasmid. HaeIII, XbaI, and HindIII have 5, 6, 14 sites, respectively.

• BMB Reports
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• 제29권1호
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• pp.52-57
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• 1996

A specific DNA fragment from Korean radish (Raphanus sativus L.) was amplified by performing PCR with oligonucleotide primers which correspond to the highly conserved regions of plant peroxidases. The size of the PCR product was ca. 400 bp, as expected from the known plant peroxidase genes. Comparison of the nucleotide and deduced amino acid sequences of the PCR product to those of other plant peroxidase-encoding genes revealed that the amplified fragment corresponded to the highly conserved region I and III of plant peroxidases. By screening a genomic library of Korean radish using the amplified fragment as a probe, two positive clones, named prxK1 and prxK2, were isolated. Restriction mapping studies indicated that the 5.2 kb Sail fragment of the prxK1 clone and the 4.0 kb EcoRI fragment of the prxK2 clone encode separate isoperoxidase genes. Analyses of the promoter region of the prxK1 clone shows that putative CAAT box, CMT box, and TGA1b binding sequence (5' TGACGT) are present 718 bp upstream from the start codon.

• 미생물학회지
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• 제24권1호
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• pp.12-17
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• 1986

A new computer algorithm is described to order the restriction fragments of plasmid DNA which has been cleaved with several restriction endonucleases in single or double digestions rapidly with realistic error rates. The permutation and high weight on small fragments methods construct all logical circular map solutions. The program is written in Apple BASIC and run on an Apple II plus microcomputer with 64K memory. Several examples are presented which indicate the high efficiency of the profram in construction possible restriction map for YEp24.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.340.1-340
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• 1995

• 미생물학회지
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• 제30권6호
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• pp.539-545
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• 1992

Lactobacillus casei SM-M1 의 플라스미드로부터 phospho-$\beta$-galactosidase gene 을 갖는 DNA 를 E. coli 에 클로닝한 pPLac15(13kb) 의 재조합 플라스미드를 제조하였다.(15). pPLac15 DNA 를 분리하여 제한효소로 처리하여 제한효소 지도를 작성하였다. Phospho-$\beta$-galactosidase 유전자의 발현을 높이기 위하여 lac promoter 를 가진 pUC18 의 PstI 위치에 클닝하여 pPLac18 을 제조하였으며, 이것을 다시 EcoRI 으로 절단하여 pUC 18 에 클로닝하여 얻은 pPLac23 (7.6 kb) 를 얻었다. Phospho-$\beta$-galactosidase 효소활성은 pPLac23 의 형질전환주인 E. coli SW-23 에서는 pPLac15 를 가진 형질전환주인 E. coli SW-15 보다 약 1.8 배의 효소의 활성을 나타내었으며 pPLac18 을 가진 E. coli SW-18 보다는 약간 높은 활성을 나타내었다.

• The Plant Pathology Journal
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• 제18권5호
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• pp.266-270
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• 2002

Inbred lines of Chinese cabbage KU-101 (resistant line against Plasmodiophora brassicae race race 6) and CS-113 (susceptible line) were crossed and their progeny lines F$_1$, BC$_1$F$_1$, F$_2$, and F$_3$ were produced for the construction of the genetic linkage map of R brassicae race 6-resistant Brassica campestris ssp. pekinensis genome. Restriction fragment length polymorphism (RFLP) was applied to compare between parents and their f$_2$ progenies with a total of 192 probes and 5 restriction enzymes. The constructed RFLP map covered 1,104 cM with a mean distance between genetic marker of 8.0 cM, and produced 10 linkage groups having 121 genetic loci. The loci of P. brassicae race 6 (CR6)-resistant Brassica genome were determined by interval mapping of quan-titative trait loci (QTL), which resulted from bioassay using the same race of the fungi in P3 population. Resistant loci were estimated in numbers 1 (Gl) and 3 (G3) linkage groups. In the regression test, Gl had a value of4.8 logarithm of odd (LOD) score, while C3 had values of 4.2-7.2. Given these results, the location of the CR6-resistant loci within the Brassica genome map can now be addressed.