• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.493-496
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• 2016

Peach rosette mosaic virus (PRMV) is a plant virus that was first reported in 1933 by Peach. It can infect hosts including peach, grape, blueberry, dandelion, plum, cherry tree, and weeds. PRMV is non-reportable in Korea, but it is designated as a controlled virus requiring plant quarantine. In this study, for the rapid and specific detection of PRMV, we developed an assay using loop-mediated isothermal amplification (LAMP). Comparison between conventional polymerase chain reaction (PCR) methods (real time-PCR and nested PCR) and LAMP for the detection of PRMV revealed an equivalent level of sensitivity by all the tested methods. For the LAMP assay, outer primer sets were used to amplify a 264-bp PCR product, which was then digested using the restriction enzyme Pvu II (CAG/CTG), and the visualization of two digestion fragments (207 + 57 bp) indicated a positive reaction. The developed LAMP assay for PRMV is expected to enable the rapid monitoring of PRMV in plants.

• The Korean Journal of Zoology
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• v.38 no.1
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• pp.87-95
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• 1995

한국산 담수어류의 잉어목, 황어아과(Leuciscinae) 어류 2속 5종의 계통적 유연관계를 구명하기 위하여 mtDNA분석을 실시하였다 6 base를 인지하는 10개의 제한효소를 처리하여 얻어진 강tDNA의 크기는 16.5-17.5 Kb였으며 Bcl I, Bgl I, Bgl II, Hin dIII, Pvu II, Xba I 등은 종간 차이가 뚜렷하였다. 각종의 집단간 mtDNA분화정도는 매우 낮았으나(p=1% 미만) M. oxyephalus의 무주집단과 제주집단은 예리적으로 큰 차이를 보였다(p=5.3%) Moroco속의 종간 분화정도를 비교한 결과 M. oxycephafus와 M. lagowsk서 사이가 평균 f=7.2%로 근면관계가 제일 가까웠고 M. keumkang과 M. semotilus는 타종들과 근연관계가 제일 멀었다. Moroco속과 Phoxinus속간의 평균 유전적 분화정도는 f=13.7%로 현저한 차이를 보였다. Brown 등(1979)의 공식을 이용하여 이들 황어아과 2속 5종의 분화시기를 추정한 결과 이들은 후기 선신세(Pliocene)와 흥적세(Pleistocene) 사이에 분화된 것으로 추정되었으며 이 결과는 동위효소 연구에서 얻어진 결파(Yang and Min, 1989)와 잘 일치한다.

• YAKHAK HOEJI
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• v.39 no.1
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• pp.6-9
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• 1995

The clinical isolate Staphylococcus aureus SA2 had four kinds of plasmids and was resistant to ampicillin, chloramphenicol, clindamycin, erythromycin, gentamicin, kanamycin, methicillin, streptomycin, tetracycline and tobramycin. Transformation experiment demonstrated that 4.44 kb plasmid(pKH6) encoded resistance to tetracycline. The cleavage map of pKH6 was determined by restriction enzyme mapping techniques. The cleavage map is given for EcoRV, HindIII, HpaI, HpaII, KpnI and Xbal. Restriction endonucleases BamHl, BglI, BGIII, BstEII, EcoRI, HaellI, PstI, PvuII, SalI, Smal, and Xhol have no site on this plasmid. The restriction map revealed extensive structural homology between pKH6 and pT181.

• Korean Journal of Microbiology
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• v.22 no.4
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• pp.249-255
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• 1984

To develop the host-vector system for industrial Coryneform bacteria that seemed to be the most suitable microorganisms for molecular breeding of genes involved in the production of amion acids, nucleotides, and other products of industrial interest, broad host range E. coli plasmid R 1162 DNA was transformed into Brevibacterium ammoniagenes and the plasmids pKU6 isolated from a transformant was physically characterized. All other plasmids from the transformed cells except pKU6 exsisted as multimeric forms in Brevibacterium ammoniagenes. The plasmid DNA was retransformed into Corynebacterium glutamicum with a high frequency ($1.32{\times}10^{-1}$ per cell) and maintained stably both in Brevibacterium ammoniagenes and Corynebacterium glutamicum after 100 generations of cultures with 25-30 copy number per cell. The size of both plasmid pKU6 and plasmid R1162 were the same and restriction maps by EcoR I, Ava I, Pst I, Pvu II and Hinc II were also similar.

• The Korean Journal of Zoology
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• v.28 no.3
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• pp.166-178
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• 1985

In order to express hepatitis B surface antigen $(HB_sAg)$ containing pre-surface antigen region in mammalian calls, 2.7 kb DNA fragment containing pre-surface region-$HB_sAg$ gene poly(A) addition site of HBV genome was cloned into simian virus 40(SV 40) based chimeric vector pSVOB. 2.7 kb DNA fragment was derived from pHBVD 107 containing tandem copies of the HBV genome in a head-to-tail arrangement by Bgl II digestion. Construction of the vector pSVOE involved the incorporation of SV40 sequences spanning the viral origin of replication and 72 bp repeats (enhancer) into a pBR 322 derivative lacking sequences which inhibit replication in mammalian cells. Bam HI linker was inserted at the Pvu II site in the proximity of SV40 late promoter of pSVOE and named as pSVOB. To construct the recombinant plasmid pSVBS, pHBVD 107 was digested with Bgl II to isolate 2.7kb DNA fragment and the fragment was ligated into the Bam HI site of pSVOB by ligation. Preliminary result showed that the recombinant plasmid pSVBS produced $HB_sAg$ in the monkey cell producing large T antigen (COS cell).

• Korean Journal of Microbiology
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• v.24 no.4
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• pp.357-363
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• 1986

Synthesis of threonine and methionine in yeast, Saccharomyces cerevisiae shares a common pathway from aspartate via homoserine. HOM6 gene encodes homoserine dehydrogenase (HSDH) which catalyzes the inter-conversion of beta-aspartate semialdehyde and homoserine. The level of HSDH is under methionine specific control. A recombinant plasmid (pEK1: 13.3kb), containing HOM6 gene, has been isolated and cloned into E. coli by complenemtary transformation of a homoserine auxotrophic yeast strain M-20-20D (hom6, trp1, ura3) to a prototrophic M20-20D/pEK1, using a library of yeast genomic DNA fragments in a yeast centromeric plasmid, YCp50(8.0kb). Isolation of HOM6has been primarily confirmed by retransformation of the original yeast strain M20-20D, using the recombinant plasmid DNA which was extracted from M20-20D/pEK1 and subsequently amplified in E. coli. Eleven cleavage sites in the insery (5.3kb) have been localized through fragment analysis for 8 restriction endonucleases; Bgl II(2 site), Bgl II(1), Cla I(3), Eco RI(1), Hind III(2), Kpn I (1), Pvu II(1) and Xho I(1).

• Korean Journal of Microbiology
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• v.35 no.1
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• pp.7-12
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• 1999

By SDS-polyacrylamide gel elcctrophoresis (SDS-PAGE) and immunoblot analysis that a protein with 66,000 daltons in size was recognized by P. aeruginosa anti-exotoxin A from P. sp. PY002. The yields of exotoxin A in P. sp. PY002 culture were influenced by the concentration of iron in the culture media. Increasing of the exotoxin A production and siderophore production was made slight increasing in the MKB medium. On the other hand, to clone the gene encoding the exoloxin A genomic library of P. sp. PY002 was constructed in pBluescript SK(+). From this library a exotoxin A homologous gene was isolated by immunological hybridization method using P. aemginosa anti-exoloxin A as a probe. Two putative clones were isolated and designated pETA23 and pETA42. Thc restriction analysis ol pETA42 demonstrated that thc 1760 bp insert contained one NcoI, PvuII, SstI, Kpnl and EcoRI site and three SmaI and HaeD sites.

• Korean Journal of Environmental Biology
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• v.21 no.1
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• pp.56-59
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• 2003

The Salmonella typhimurium cdd gene encoding cytidine deaminase (cyti-dine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5.) was isolated through shotgun clon-ing by complementation of the E. coli odd mutation. By subsequent deletion and sub-cloning from the original 3.7 Kb of EcoRI insert (pSAMI), the precise region of the cdd structural gene is located around the BglII site in the middle part of 1.7 Kb of NruI/PvuI segment. The 1.7 Kb containing odd gene wag subcloned to the pUC18 vector and the nucleotide sequence of the cdd gene was determined. When the putative ribosorne-binding site (Shine-Dalgarno sequence) and initiation codon were predicted to be GAGG at the position 459 and ATG at the position 470, respectively, there was an open reading frame of 885 nucleotides, encoding an 294 amino acid protein. The cdd gene expression in E. coli JF611/pSAMI was amplified about 50 fold compared to that of the wild type. The cdd gene expression was maintained in the stationary phase after rea-ching the peak in the late logarithmic phase.

• YAKHAK HOEJI
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• v.32 no.4
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• pp.213-221
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• 1988

Inducible MLS resistance gene of Bacillus licheniformis specified by erm K was subcloned in Bacillus subtilis and the DNA sequence corresponding to its control region was determined. The determinant erm K was in Pvu II=Hind III fragment, which was 1.3 kb. The leader region is capable of forming a complex series of inverted complementary repeat sequences (ICRS) centering on at least six axes of symmetry, some of them mutually exclusive, in a way that resulted ultimately in post-transcriptional unmasking of the ribosome loading site for methylase synthesis.

• Animal Systematics, Evolution and Diversity
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• v.13 no.2
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• pp.73-81
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• 1997

한국산 조류 중 박새속(genus Parus)에 속하는 4종 Parus major wladiwostokensis (박새), P. ater amurensis(진박새), P. palustrius hellmayri(쇠박새) 및 P. varius varius(곤줄 박이)를 대상으로 이들의 계통적 유연관계를 구명하기 위하여 mtDNA분석을 실시하였다. 6 base를 인지하는 10개의 제한효소를 처리하여 얻어진 mtDNA의 크기는 16.6~17.0Kb였으며 Pst I과 Pvu II는 종간 차이가 뚜렷하였다. 각 종간의 절편양상을 비교하여 Parus속의 종 간 분화정도를 비교한 결과 P. m. wladiwostokensis와 P. a. amurensis의 유전적 근연관계 가 가장 가까웠고(p=0.073, F=0.294) P. a. amurensis와 P. v. varius는 비교적 현저한 유전 적 차이를 보였다.(p=0.119, F=0.143). Brown 등 (1979)의 공식을 이용하여 박새속 4종의 분 화시기를 추정한 결과 이들은 후기 선신세(Pliocene)와 초기 홍적세(Pleistocene) 사이에 분 화된 것으로 추정되었다.