• 미생물학회지
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• 제33권3호
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• pp.165-169
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• 1997

병원하수에서 분리한 gentamicin 저항성 세균으로부터 aacC2 유전자를 가지고 있는 R 플라스미드 pGM5와 pGM6를 선발하였다. 이들 플라스미드에서 gentamicin 저항성 유전자를 포함하는 부분을 pUC18의 BamHI 자리에 클로닝하여 재조합 플라스미드 pSY5와 pSY6를 각각 얻었다. 재조합 플라스미드의 삽입된 부분에 대한 제한효소 지도를 통해 Tn3 염기서열이 aacC2 유전자의 상류부위에 위치하는 것을 알았다. 재조합 플라스미드의 gentamicin에 대한 민감성의 비교를 통해 Tn3의 bla 유전자 부분과 3'역행중복 부위의 염기서열이 gentamicin 저항성 유전자의 발현에 중요한 역할을 담당하고 있었다.

• 식물분류학회지
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• 제51권3호
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• pp.326-331
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• 2021

The complete chloroplast genome of Zoysia macrostachya Franch. & Sav. isolated in Korea is 135,902 bp long (GC ratio is 38.4%) and has four subregions; 81,546 bp of large single-copy (36.3%) and 12,586 bp of small single-copy (32.7%) regions are separated by 20,885 bp of inverted repeat (44.1%) regions, including 130 genes (83 protein-coding genes, eight rRNAs, and 39 tRNAs). Thirty-nine single nucleotide polymorphisms and 11 insertions and deletion (INDEL) regions were identified from two Z. macrostachya chloroplast genomes, the smallest among other Zoysia species. Phylogenetic trees show that two Z. macrostachya chloroplast genomes are clustered into a single clade. However, we found some incongruency with regard to the phylogenetic position of the Z. macrostachya clade. Our chloroplast genome provides insights into intraspecific variations and species delimitation issues pertaining to the Zoysia species.

• Journal of Species Research
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• 제10권2호
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• pp.154-158
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• 2021

An endemic endangered species, Aster altaicus var. uchiyamae (Danyang aster) B2015-0044, is cultivated at the Shingu Botanical Garden, which serves as the ex situ conservation institution for this species. In this work, we sequenced the chloroplast genome of A. altaicus var. uchiyamae B2015-0044. We found that the chloroplast (cp) genome of B2015-0044 was 152,457 base pairs(bps) in size: 84,247 bps of large single copy regions(LSC), 25,007 bps of inverted repeats(IRs), and 18,196 bps of small single copy regions. The B2015-0044 cp genome contains 79 protein-coding genes (PCGs), 4 RNA genes, 29 tRNA genes, and 3 pseudogenes. These results were identical to a previously reported cp genome (Park et al., 2017), except for two sites in introns and three in intergenic spacer (IGS) regions. For the intronic differences, we found that clpP.i1 had a 1-bp small simple repeat (SSR) (T) and petD.i had a 3-bp SSR (ATT). We found 1-bp SSRs in the IGSs of trnT_ggu~psbD and psbZ~trnG_gcc, C and A, respectively. The IGS of(ndhF)~rpl32 had a SNP. Based on our results, the cp genome of the A. altaicus var. uchiyamae can be classified into two genotypes, [C]¹-[A]¹²-[T]¹²-[ATT]⁴-C and [C]²-[A]¹¹-[T]¹¹-[ATT]²-A.

• 식물분류학회지
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• 제51권1호
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• pp.109-114
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• 2021

Veronica nakaiana Ohwi (Plantaginaceae) is an endemic taxon on Ulleungdo Island, Korea. We report the second complete chloroplast genome sequence of V. nakaiana. Its genome size is 152,319 bp in length, comprising a large single-copy of 83,195 bp, a small single-copy of 17,702 bp, and a pair of inverted repeat regions of 25,711 bp. The complete genome contains 115 genes, including 51 protein-coding genes, four rRNA genes, and 31 tRNA genes. When comparing the two chloroplast genomes of V. nakaiana, 11 variable sites are recognized: seven SNPs and four indels. Two substitutions in the coding regions are recognized: rpoC2 (synonymous substitution) and rpl22 (nonsynonymous substitution). In nine noncoding regions, one is in the tRNA gene (trnK-UUU), one is in the intron of atpF, and seven are in the intergenic spacers (trnH-GUG~psbA, trnK-UUU, rps16~trnQ-UUG, trnC-GCA~petN, psbZ~trnG-GCC, ycf3~trnS-GGA, ycf4~cemA, and psbB~psbT). The data provide the level of genetic variation in V. nakaiana. This result will be a useful resource to formulate conservation strategies for V. nakaiana, which is a rare endemic species in Korea.

• 식물분류학회지
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• 제51권4호
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• pp.345-352
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• 2021

Diarthron linifolium Turcz. is an annual herb usually found in sandy soil or limestone areas. Plants in the genus Diarthron are known to have toxic chemicals that may, however, be potentially useful as an anticancer treatment. Diarthron linifolium is a unique species among the species of the genus distributed in Korea. Here, we determine the genetic variation of D. linifolium collected in Korea with a full chloroplast genome and investigate its evolutionary status by means of a phylogenetic analysis. The chloroplast genome of Korean D. linifolium has a total length of 172,644 bp with four subregions; 86,158 bp of large single copy and 2,858 bp of small single copy (SSC) regions are separated by 41,814 bp of inverted repeat (IR) regions. We found that the SSC region of D. linifolium is considerably short but that IRs are relatively long in comparison with other chloroplast genomes. Various simple sequence repeats were identified, and our nucleotide diversity analysis suggested potential marker regions near ndhF. The phylogenetic analysis indicated that D. linifolium from Korea is a sister to the group of Daphne species.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.55-55
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• 2022

Corylopsis Siebold & Zucc. (Hamamelidaceae) is widely used for horticultural plant and comprise ca. 25 species in East Asia (1 species in Korea; 4 species in Japan; 20 species in China). Previous revisions have gone from 7 to more than 30 species, causing confusion in the nursery industry and public gardens. Due to morphological similarity within Corylopsis, molecular research is needed to distinguish it. In this study, the chloroplast genome of C. gotoana and C. pauciflora distributed in Japan was completed by using NGS (Next-Generation Sequencing) technique. The genome size of C. gotoana and C. pauciflora were 159,434 bp (large single-copy (LSC): 88,164 bp; small single-copy (SSC): 18,702 bp; inverted repeat regions (IRs): 26,284 bp) and 159,363 bp (LSC: 88,097 bp; SSC: 18,700 bp; IRs: 26,283 bp), respectively. In addition, we investigated the repeats, SNPs, and indels, and that could be used as DNA markers. Phylogenetic analysis demonstrated that C. pauciflora was sister to C. gotoana and C. spicata. The genus Corylopsis is a monophyletic group and Loropetalum is closely related to Corylopsis. The results of our study will provide the basic data necessary for the analysis of the species identification markers and genetic diversity within the genus Corylopsis in the future.

• Molecules and Cells
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• 제26권5호
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• pp.496-502
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• 2008

Cryparin, encoded as a single copy gene (Crp) of the chestnut blight fungus Cryphonectria parasitica, is the most abundant protein produced by this fungus. However, its accumulation is decreased remarkably in C. parastica strains containing the double-stranded (ds) RNA virus Cryphonectria hypovirus 1. To characterize the transcriptional regulatory element(s) for strong expression and viral regulation, promoter analysis was conducted. Serial deletion of the Crp promoter region resulted in a step-wise decrease in promoter activity, indicating a localized distribution of genetic elements in the cryparin promoter. Promoter analysis indicated two positive and a repressive cis-acting elements. Among them, the promoter region between nt -1,282 and -907 appeared to be necessary for hypoviral-mediated down-regulation. An electrophoretic mobility shift assay (EMSA) on the corresponding promoter region (-1,282/-907) indicated two regions at (-1,257/-1,158) and (-1,107/-1,008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.

• 식물병연구
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• 제18권4호
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• pp.268-276
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• 2012

Broad bean wilt virus 2 (BBWV2), genus Fabavirus, subfamily Comovirinae, family Secoviridae, causes damage in many economically important horticultural and ornamental crops. Sequence alignments showed several conserved sequences in 5' non-coding regions (5' NCRs) of RNA 1 and RNA 2 in all BBWV2 strains characterized so far. Based on this observation, we generated a hpRNA construct (pIR-BBWV2) harboring an inverted repeat containing a 210 bp cDNA fragment homologous to 5' NCR portion of BBWV2 RNA 1 to investigate the silencing potential for its ability to interfere with a rapidly replicating BBWV2. Agrobacterium-mediated transient expression of the IR-BBWV2 had a detrimental effect on BBWV2 infection, showing no distinct symptoms in non-inoculated leaves of the agroinfiltrated Nicotiana benthamiana plants. BBWV2 genomic RNAs were not detected by RT-PCR from tissues of both the inoculated leaves and upper leaves of the agroinfiltrated plants. Accumulation of virus-derived small interfering RNAs was detected in the inoculated leaf tissues of N. benthamiana plants elicited by transient expression of IR-BBWV2 indicating that RNA silencing is responsible for the resistance to BBWV2.

• 식물조직배양학회지
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• 제22권5호
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• pp.241-260
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• 1995

Transposons, present in the genomes of all living organisms, are genetic element that can change positions, or transpose, within the genome. Most genomes contain several kinds of transposable elements and the molecular details of the mechanisms by which these transposons move have recently been uncovered in many families of transposable elements. Transposition is brought about by an enzyme known as transposaese encoded by the autonomous transposon itself, but, in the unautonomous transposon lacking the gene encoding the transposase, movement occurs only at the presence of the enzyme encoded by the autonomous one. There are two types of transposition events, conservative and replicative transposition. In the former the transposon moves without replication, both strands of the DNA moving together from one place to the other while in the latter the transposition frequently involves DNA replication, so one copy of transposon remains at its original site as another copy insole to a new site. The insertion of transposon into a gene can prevent it expression whereas excision from the gene may restore the ability of the gene to be expressed. There are marked similarities between transposons and certain viruses having single stranded Plus (+) RNA genomes. Retrotransposons, which differ from the ordinary transposons in that they transpose via an RNA-intermediate, behave much like retroviruses and have a structure of integrated retrovial DNA when they are inserted to a new target site. An insertional mutagenesis called transposon-tagging is now being used in a number of plant species to isolate genes involved in developmental and metabolic processes which have been proven difficult to approach by the traditional methods. Attempts to device a transposon-tagging system based on the maize Ac for use in heterologous species have been made by many research workers.

• Journal of Microbiology and Biotechnology
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• 제18권8호
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• pp.1357-1363
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• 2008

The $P_{entC}$ promoter of the entCERA operon encoding enzymes for enterobactin biosynthesis in Escherichia coli is tightly regulated by the availability of iron in the culture medium. In iron-rich conditions, the $P_{entC}$ promoter activity is strongly repressed by the global transcription regulator Fur (ferric uptake regulator), which complexes with ferrous ions and binds to the Fur box 19-bp inverted repeat. In this study, we have constructed the expression vector pOS2 containing the $P_{entC}$ promoter and characterized its repression, induction, and modulation by quantifying the expression of the lacZ reporter gene encoding $\beta$-galactosidase. $\beta$-Galactosidase activities of E. coli transformants harboring pOS2-lacZ were highly induced in the presence of divalent metal ion chelators such as 2,2'-dipyridyl and EDTA, and were strongly repressed in the presence of excess iron. It was also shown that the basal level $\beta$-galactosidase expression by the $P_{entC}$ promoter was drastically decreased by incorporating the fur gene into the expression vector. Since the newly developed iron chelator-inducible expression system is efficient and cost-effective, it has wide applications in recombinant protein production.