• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1106-1112
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• 2007

This report provides the complete nucleotide sequences of the full-length cDNA encoding squalene synthase (SQS) and its genomic DNA sequence from a triterpene-producing fungus, Ganoderma lucidum. The cDNA of the squalene synthase (SQS) (GenBank Accession Number: DQ494674) was found to contain an open reading frame (ORF) of 1,404 bp encoding a 468-amino-acid polypeptide, whereas the SQS genomic DNA sequence (GenBank Accession Number: DQ494675) consisted of 1,984 bp and contained four exons and three introns. Only one gene copy was present in the G. lucidum genome. The deduced amino acid sequence of Ganoderma lucidum squalene synthase (GI-SQS) exhibited a high homology with other fungal squalene synthase genes and contained six conserved domains. A phylogenetic analysis revealed that G. lucidum SQS belonged to the fungi SQS group, and was more closely related to the SQS of U. maydis than to those of other fungi. A gene expression analysis showed that the expression level was relatively low in mycelia incubated for 12 days, increased after 14 to 20 days of incubation, and reached a relatively high level in the mushroom primordia. Functional complementation of GI-SQS in a SQS-deficient strain of Saccharomyces cerevisiae confirmed that the cloned cDNA encoded a squalene synthase.

• Animal cells and systems
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• 제9권2호
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• pp.87-94
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• 2005

Spindle position checkpoint monitors the orientation of mitotic spindle for proper segregation of replicated chromosomes into mother cell and the daughter, and prohibits mitotic exit when mitotic spindle is misaligned. BUB2 forms one of the key upstream element of spindle position checkpoint in budding yeast, but its functional homologues have not been identified in higher eukaryotes. Here, we analyzed the functions of two putative BUB2 homologues of C. elegans in the spindle orientation checkpoint. From the C. elegans genome database, we found that two open reading frames (ORFs), F35H12_2 and C33F10_2, showed high sequence homology with BUB2. We obtained the expressed sequence tag (EST) clones for F35H12_2 (yk221d4) and C33F10_2 (yk14e10) and verified the full cDNA for each ORF by sequencing and 5' RACE with SL1 primer. The functional complementation assays of yk221d4 and yk14e10 in ${\Delta}bub2$ of S. cerevisiae revealed that these putative BUB2 homologues of C. elegans could not replace the function of BUB2 in spindle position checkpoint and mitotic exit. Our attempt to document the component of spindle position checkpoint in metazoans using sequence homology was not successful. This suggests that structural information about its components might be required to identify functional homologues of the spindle position checkpoint in higher eukaryotes.

• BMB Reports
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• 제33권3호
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• pp.195-201
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• 2000

A human gene has been reported that may encode the enzyme acetohydroxyacid synthase. Previously this enzyme was thought to be absent from animals although it is present in plants and many microorganisms. In plants, this enzyme is the target of a number of commercial herbicides and the use of these compounds may need to be reassessed if the human enzyme exists and proves to be susceptible to inhibition. Here we report the construction of several plasmid vectors containing the cDNA sequence for this protein, and their expression in Escherichia coli. High levels of expression were observed, but most of the protein proved to be insoluble. The small amounts of soluble protein contained little or no acetohydroxyacid synthase activity. Attempts to refold the insoluble protein were successful insofar as the protein became soluble. However, the refolded protein did not gain any acetohydroxyacid synthase activity. In vivo complementation tests of an E. coli mutant produced no evidence that the protein is active. Incorrect folding, or the lack of another subunit, may explain the data but we favor the interpretation that this gene does not encode an acetohydroxyacid synthase.

• BMB Reports
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• 제40권3호
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• pp.349-357
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• 2007

The ubiquitous family of 14-3-3 proteins functions as regulators in a variety of physiological processes. Eight rice 14-3-3 genes, designated OsGF14a through h, were identified from an exhaustive search of the genome database. Comparisons of deduced amino acid sequences reveal a high degree of identity among members of the OsGF14 family and reported Arabidopsis 14-3-3 proteins. A phylogenetic study indicates that OsGF14s contain both $\varepsilon$ and non-$\varepsilon$ forms, which is also confirmed by a structural analysis of OsGF14 genes. Furthermore, transcripts of OsGF14b, OsGF14c, OsGF14d, OsGF14e, OsGF14f and OsGF14g were detected in rice tissues. Their different expression patterns, the different effects of environmental stresses and plant hormones on their transcription levels, and the different complementary phenotypes in yeast 14-3-3 mutants not only indicates that OsGF14s are responsive to various stress conditions and regulated by multiple signaling pathways, but also suggests that functional similarity and diversity coexist among the members of OsGF14 family.

• 한국환경과학회지
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• 제17권8호
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• pp.899-905
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• 2008

Quality test for activated carbon(AC) filter used for drinking water purifier is now an obligatory test and the standard material for valid purifying amount in water purifier performance test has been changed from residual chlorine to chloroform according to the notice of Ministry of Environment in 2006. Therefore, this study aimed to compare the ingredients of AC filters by confirming chloroform removal rate of AC filter and conducting 4 adsorption tests (Iodine, methylene blue decolorization, phenol value, ADS value) for AC filters provided by manufacturers. With water pressure of $1kgf/cm^2$, 1,500 liters of prepared inflow went through to check chloroform removal rate. As a result, product with removal rate of below 60% from all products. On the other hand, 4 adsorption tests were conducted for filters in the market and filters from manufacturers. None of the products satisfied all 4 tests. In particular, they showed great shortage to the standard in phenol value and ADS value test. However, manufacturers' filter showed much better performance than filters in the market. Also, the result of valid purifying amount test for each of five products of appropriate product and inappropriate product based on filter quality test showed average 4,440 liters for appropriate product and average 2,620 liters for inappropriate product. According to the result, it is shown that the filter with good adsorption also had good chloroform removal efficiency and adsorption efficiency. Therefore, it is expected that customers can screen good quality product through obligatory conduct of filter quality test. However, it is considered that complementation in system is required for future inspection.

• 미생물학회지
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• 제50권3호
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• pp.249-253
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• 2014

THOC7/Mft1는 mRNA가 전사되는 동안 mRNP의 포장과 mRNA 방출에 관여하는 진화적으로 잘 보존된 THO 복합체의 구성인자이다. 분열효모 Schizosaccharomyces pombe에서 THOC7/Mft1의 이종상동체(spThoc7)가 합성치사 돌연변이체 SLRsm1의 생장 결함을 부분적으로 상보하는 것으로 선별되었다. 이배체 S. pombe 균주에 하나의 spthoc7 유전자만을 결실시킨 후 4분체 분석을 수행한 결과, 이 유전자는 생장에 필수적이지 않았다. 하지만, ${\Delta}thoc7$ 결실돌연변이는 생장과 mRNA의 핵에서 세포질로의 방출에 약간의 결함을 보였다. 기능을 하는 spThoc7-GFP단백질은 주로 핵 안에 존재하였다. 이와 같은 결과들은 spThoc7도 mRNA 방출에 관여하고 있음을 시사한다.

• 미생물학회지
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• 제37권1호
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• pp.42-48
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• 2001

출아 효모인 Saccharomyces cerevisiae의 베타-1,3-글루칸 생합성의 결함을 초래하는 돌연변이 유전자(sool-1)를 분리하여 돌연변이 부위의 염기서열을 결정하고 그 특성을 분석하였다. cool-1 유전자의 염기서열 분석 결과, 681번의 염기인 G가 A로 치환되어 Soolp의 $Gly^{227}$이 Asp로 치환되는 결과를 나타내는 것으로 판명되었고, cool-1 유전자는 기존에 보고된 retl-1 유전자와 동일한 돌연변이 유전자로 판명되었다. 그러나, ret1-1이 나타내는 온도감수성 형질은 배지에 1.2 M sorbitol 등의 삼투안정제를 첨가하면 극복될 수 있으며, cool-l/retl-1의 돌연변이 부위가 세포벽합성 관련 단백질의 번역 후 수식과정에 영향을 미친 것임을 확인하였다. 한편, Soolp/$\alpha$-COP의 N-말단에 존재하는 6개의 WD40 domain중 5번째 WD40 domain이 효모의 세포벽 합성이나 구조유지에 중요한 역할을 담당할 것임을 시사하는 결과를 얻었다.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.28-28
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• 2015

Asexual reproduction or conidiation in aspergilli is a primary mean to produce their progenies that is environmentally and genetically controlled tightly. Previously, intensive researches in the model fungus Aspergillus nidulans disclosed some genes playing important roles in asexual and sexual development. Among them, one gene encoding a putative helix-loop-helix (HLH) transcription factor, named ndrA, has been isolated and characterized as a downstream regulator of developmental master regulator NsdD. By using comparative genome search of A. niduans NdrA protein, its orthologues have been identified in A. fumigatus and A. flavus, respectively (AfudrnA and AfldrnA). Deletion of the ndrA genes in both Aspergillus species made them unable to produce the conidia yet abundant production of sclerotia in A. flavus. Complementation of ndrA deletion strains by intact ndrA ORFs has restored the conidiation as in the control strains. In A. fumigatus, ndrA deletion also resulted in loss of conidiation phenotype. Northern analyses showed that the ndrA genes in both Aspergillus species are highly expressed at the early stage of the conidiation. Interestingly, the ndrA genes were found to be necessary for the proper expression of brlA genes. Antifungal sensitivity test revealed that the ndrA genes might be responsible for the sensitivity or resistance to some antifungal agents. However, ndrA deletion did not greatly influence the growth in both strains. And the A. flavus ndrA gene did not affect the aflatoxin production. Taken together, ndrA genes in Aspergillus species could be an important positive regulator of conidiation under the regulation of the nsdD gene yet upstream of the brlA gene.

• Journal of Microbiology and Biotechnology
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• 제21권9호
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• pp.903-913
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• 2011

Vibrio parahaemolyticus, which causes gastroenteritis, wound infection, and septicemia, has two sets of type III secretion systems (TTSS), TTSS1 and TTSS2. A TTSS1-deficient vcrD1 mutant of V. parahaemolyticus showed an attenuated cytotoxicity against HEp-2 cells, and a significant reduction in mouse lethality, which were both restored by complementation with the intact vcrD1 gene. V. parahaemolyticus also triggered phosphorylation of mitogen-activated protein kinases (MAPKs) including p38 and ERK1/2 in HEp-2 cells. The ability to activate p38 and ERK1/2 was significantly affected in a TTSS1-deficient vcrD1 mutant. Experiments using MAPK inhibitors showed that p38 and ERK1/2 MAPKs are involved in V. parahaemolyticus-induced death of HEp-2 cells. In addition, caspase-3 and caspase-9 were processed into active forms in V. parahaemolyticus-exposed HEp-2 cells, but activation of caspases was not essential for V. parahaemolyticus-induced death of HEp-2 cells, as shown by both annexin V staining and lactate dehydrogenase release assays. We conclude that secreted protein(s) of TTSS1 play an important role in activation of p38 and ERK1/2 in HEp-2 cells that eventually leads to cell death via a caspase-independent mechanism.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.582-588
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• 1999

A Brevibacterium lactofermentum gene coding for a glucose-specific permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned, by complementing an Escherichia coli mutation affecting a ptsG gene with the B. lactofermentum genomic library, and completely sequenced. The gene was identified as a ptsG, which enables an E. coli transformant to transport non-metabolizable glucose analogue 2-deoxyglucose (2DG). The ptsG gene of B. lactofermentum consists of an open reading frame of 2,025 nucleotides encoding a polypeptide of 674 amino acid residues and a TAA stop codon. The 3' flanking region contains two stem-loop structures which may be involved in transcriptional termination. The deduced amino acid sequence of the B. lactofermentum enzyme $II^{GIe}$ specific to glucose ($EII^{GIe}$) has a high homology with the Corynebacterium glutamicum enzyme $II^{Man}$ specific to glucose and mannose ($EII^{Man}$), and the Brevibacterium ammoniagenes enzyme $II^{GIc}$ specific to glucose ($EII^{GIc}$). The 171-amino-acid C-terminal sequence of the $EII^{Glc}$ is also similar to the Escherichia coli enzyme $IIA^{GIc}$ specific to glucose ($IIA^{GIc}$). It is interesting that the arrangement of the structural domains, IIBCA, of the B. lactofermentum $EII^{GIc}$ protein is identical to that of EIIs specific to sucrose or $\beta$-glucoside. Several in vivo complementation studies indicated that the B. lactofermentum $EII^{Glc}$ protein could replace both $EII^{ Glc}$ and $EIIA^{Glc}$ in an E. coli ptsG mutant or crr mutant, respectively.