• Journal of Microbiology
• /
• v.34 no.3
• /
• pp.248-254
• /
• 1996

$cdc103^+$ gene in Schizosaccharomyces pombe which is similar to the CDC3 gene in Saccharomyces cerevisiae was cloned and sequenced. Comparison of the predicted amino acid sequences of $cdc103^+$ and CDC3 revealed that they share significant similarity (43% identity and 56% identity or similarity) to each other. The gene product of CDC3 in S. cerevisiae is known to be a highly ordered ring of filaments that lies just inside the cytoplasmic membrane in the region of the mother-bud neck. In order to characterize the gene product of $cdc103^+$ in Schizosaccharomyces pombe, fusion proteins were used to generate the polyclonal antibodies specific for the gene product (cdc103p). In immunofluorescence experiments, these antibodies decorate the region of the septum formation as a double ring structure late in the cell division cycle.

• Microbiology and Biotechnology Letters
• /
• v.24 no.4
• /
• pp.415-422
• /
• 1996

A gene coding for a pyruvyl transferase enzyme involved in exopolysaccharide biosynthesis of Zoogloea ramigera 115SLR was isolated and sequenced. A 4.5 kb of BamHI DNA fragment was isolated from chromosomal DNA using a probe derived from ketal pyruvyl transferase gene of Xanthomonas campestris. The nucleotide sequence of 2.66 kb Pst1/HindIII DNA fragment which was homology with a probe revealed the existence of two complete open reading frames (ORF2 and ORF3) and two partial open reading frames (ORFI and ORF4). The deduced amino acid sequence of ORF3 was homologous to the ketalase (GumL product) of X campestris with 49.5% of similarity and 21.6% of identity. ORF2 on the other hand showed the higher identity with the ketalase (ExoV product) of Rhizobium meliloti (36%) as well as the ketalase of X campestris (23%) than that of ORF3. A gene product of ORF2 was determined with a bacteriophage T7 RNA polymerase/promoter system in E. coli. The molecular weight of protein was 33,500 dalton.

• The Microorganisms and Industry
• /
• v.17 no.1
• /
• pp.2-9
• /
• 1991

All of four genes are cloned and DNA sequence analysis have now revealed that these four genes encode a family of proteins with similar amino acid sequence. These proteins show no extensive similarities to any known proteins (Haarer et al., 1991). Among them, CDC3 gene is fused with E. coli lacZ and trpE genes and antibodies against the CDC3 gene product are produced. These antibodies are used to check the localization of this product to the vicinity of the 10 -nm filaments in the mother-bud neck.

• Journal of Life Science
• /
• v.8 no.6
• /
• pp.673-680
• /
• 1998

Adenosine deaminase gene from Nocardioides sp. J-326TK was cloned by polymerase chain reaction using primers (PI, PII and PIII) constructed from the highly conserved amino acid sequences among Escherichia coli, mouse and human. A PCR product of about 800bp, as expected from the sequence of E. coli adenosine deaminase gene, was obtained from Nocardioides sp. J-326TK chromosomal DNA double-digested with EcoRI and Pst I. DNA sequencing of the PCR product after cloning into pT7Blue T-vector shows 99.5% and 98.9% homologies in nucleotide and amino acid sequences, respectively, with the E. coli adenosine deaminase whereas 59.5% and 46.8% homologies with the human adenosine deaminase, indicating the evolutionarily relationship of these organisms.

• The Microorganisms and Industry
• /
• v.19 no.4
• /
• pp.32-35
• /
• 1993

ras는 활성화 형태인 GTP bound form과 비활성화 형태인 GDP bound form의 두 형태로 존재하며 두 형태를 매개하는 regulatory protein들에 의해 그 activity가 조절된다. 또한 ras는 GTP와 GDP에 강한 친화성이 있으며 세포내에는 GTP보다 GDP가 더 많이 있어서 평소에는 ras가 GDP와 결합하고 있다가 활성화될때만 GTP와 결합하는 것으로 추정된다. GDP bound ras는 guanine nucloetide exchange protein(GEP)에 의해 활성화된 GTP bound form으로 전환되며 ras의 기능이 발휘된 후에는 GTPase activating protein(GAP)에 의해 비활성화된다. Yeast의 경우 IRA1과 2의 product가 GAP의 역할을 하는 것으로 알려져 있고 CDC25 gene의 product가 GEP의 기능을 담당하는 것으로 알려져 있다. NF1 gene은 Von Recklinghausen Neurofibromatosis Type I 질병을 가진 환자에게서 발견되었는데 부분적으로 sequencing한 결과에 따르면 yeast의 IRA1/2, mammalian GAP gene product와 protein homology가 높은 것으로 나타났다. Yeast의 경우 IRA1/2 gene의 손실이나 mammalian ras gene의 transformation으로 인한 heat shock sensitivity가 NF1 gene(2,3) 혹은 GAP(4)의 expression으로 suppression된 것으로 보아 NF1이 GAP protein으로서 ras를 불활성화 시킨다는 것이 판명되었다. 결론적으로 ras의 활성은 GTP bound 혹은 GDP bound의 양쪽형태를 이동하면서 조절되는데 이 기능은 GAP과 GEP 또는 그의 유사 protein들에 의해 수행되며 이러한 regulatory protein들은 growth factor, cytokine 그리고 protein kinase 같은 signal에 의해 활성화된다고 생각된다. 본 총설에서는 ras protein의 여러가지 성질보다는 ras의 modification과 관련하여 항암제로 사용할 수 있는 ras에 specific한 약품개발의 가능성과 현재 알려진 ras의 inhibitor를 중심으로 논하고자 한다.

• Journal of Microbiology and Biotechnology
• /
• v.6 no.5
• /
• pp.331-335
• /
• 1996

The second $\beta$-Xylosidase gene (xylB) from Bacillus stearothermophilus was isolated from the genomic library, cloned into pBR322, and subsequently transferred into Escherichia coli HB101. Six out of 10, 000 transformants were selected from the selective LB medium supplemented with p-nitrophenyl-$\alpha$-L-arabinofuranoside (pNPAf) and ampicillin ($50\mu g$/ml) based on their ability to form a yellow ring around the colony. One of the clones was found to harbor the recombinant plasmid with 5.0 kb foreign DNA, which was identical to the $\alpha$-L-arabinofuranosidase gene (arfI) previously cloned in this lab, while the other five had 3.5 kb of the foreign DNA. Southern blotting experiments confirmed that the 3.5 kb insert DNA was from B. stearothermophilus chromosomal DNA. A zymogram with 4-methylumbelliferyl-$\alpha$-L-arabinofuranoside as the enzyme substrate revealed that the cloned gene product was one of the mutiple $\alpha$-L-arabinofuranosidases produced by B. stearothermophilus. Unlike the arfI gene product, the product of the gene on the insert DNA (xylB) showed an activity not only on pNPAf but also on oNPX suggesting that the cloned gene product could be a bifunctional enzyme having both $\alpha$-L-arabinofuranosidase and $\beta$-xylosidase activities.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.7
• /
• pp.1276-1280
• /
• 2017

The mcr-1 gene is a new "superbug" gene discoverd in China in 2016 that makes bacteria highly resistant to the last-resort class of antibiotics. The mcr-1 gene raised serious concern about its possible global dissemination and spread. Here, we report a potential anti-resistant strategy using the CRISPR/Cas9-mediated approach that can efficiently induce mcr-1 gene knockout in Escherichia coli. Our findings suggested that using the CRISPR/Cas9 system to knock out the resistance gene mcr-1 might be a potential anti-resistant strategy. Bovine myeloid antimicrobial peptide-27 could help deliver plasmid pCas::mcr targeting specific DNA sequences of the mcr-1 gene into microbial populations.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.10
• /
• pp.1648-1652
• /
• 2015

Azaphilone polyketides are synthesized by iterative non-reducing fungal polyketide synthases (NR-fPKSs) with a C-terminus reductive domain (-R). Several azaphilone biosynthetic gene clusters contain a putative serine hydrolase gene; the Monascus azaphilone pigment (MAzP) gene cluster harbors mppD. The MAzP productivity was significantly reduced by a knockout of mppD, and the MAzP NR-fPKS-R gene (MpPKS5) generated its product in yeast only when co-expressed with mppD. Site-directed mutations of mppD for conserved Ser/Asp/His residues abolished the product formation from the MpPKS5/mppD co-expression. MppD and its homologs are thus proposed as a new protein factor involved in the product formation of NR-fPKS-R.

• Journal of Microbiology
• /
• v.38 no.3
• /
• pp.156-159
• /
• 2000

The cttl+ gene in Schizosaccharomyces pombe encoeds a catalse responsible for H2O2-resistance of this organism as judged by the H2O2-sensitive phenotype of the ctt1Δ mutant. In this study, we investigated the subcellular localization of the Ctt1 gene product. In wild type cells catalase activity was detected in the organelle fraction as well as in the cytosol. The ctt1Δ mutant contained no catalase activity, indicating that both cytosolic and organellar catalases are the products of a single ctt1+ gene. Western bolt analysis revealed two catalase bands, both of which disappeared in the ctt1Δ mutant. The major, fastermigrating band existed in the cytosol whereas the monor, slower-migrating band appeared to be located in organelles, most likely in peroxisomes. These results suggest that the ctt1+ gene product targeted to the peroxisome is a modified form of the one in the cytosol.

• Proceedings of the Zoological Society Korea Conference
• /
• 2001.10a
• /
• pp.86-86
• /
• 2001

No Abstract, See Full Text