• Title/Summary/Keyword: ${\beta}-Tubulin$ gene

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Bombyx mori β-tubulin Promoter for High-level Expression of Heterologous Genes

  • Park, Kwanho;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.39 no.1
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    • pp.22-28
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    • 2019
  • We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with ${\beta}$-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5'-flanking region of ${\beta}$-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the ${\beta}$-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that ${\beta}$-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.

Cloning of $\beta$-Tubulin Gene and Effect of Pencycuron on Tubulin Assembly in Rhizoctonia solani

  • Roh, Seong-Hwan;Heung-Tae KIm;Isamu-Yamaguchi
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.68-71
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    • 1999
  • To illustrate the action mechanism of pencycuron on Rhizoctonia solani, two experiments were conducted including the comparison of amino acids of $\beta$-tubulin between R-C (sensitive isolate) and Rh-131 (non-sensitive isolate), and the inhibitory effect of pencycuron on tubulin assembly in vitro. Both $\beta$-tubulin genes of R-C and Rh-131 proved to have 1,582 nucleotides encoding a protein of 445 amino acids, showing 98% homology in amino acid sequences between them. It was found that codons at 103, 236, and 267 for lysine (AGG), valine (GTC) and isoleucine (ATT) in R-C were replaced by codons for methionine (ATG), isoleucine (ATT) and methionine (ATG) in Rh-131, respectively. No inhibitory effect of pencycuron on the tubulin assembly was observed. It suggests that pencycuron may have no direct inhibitory effects on the assembly of tubulin at least in vitro.

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Amino Acid Alterations in the $\beta$- Tubulin of Metarhizium anisopliae That Confer Benomyl Resistance

  • Kim Soon Kee;Shim Hee Jin;Roh Jong Yul;Li Ming Shun;Choi Jae Young;Jin Byung Rae;Boo Kyung Saeng;Je Yeon Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.1
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    • pp.37-42
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    • 2005
  • We cloned the $\beta$-tubulin genes from the wild type strain and two benomyl-resistant mutants of Metahizium anisopliae and determined their nucleotide sequences. A $\beta$-tubulin encoding 448-residue protein from wild type M. anisopliae shows strong homology to other $\beta$-tubulins. The coding region is interrupted by four introns. Comparisons of intron position between the M. anisopliae gene and other fungal $\beta$-tubulin genes show considerable positional conservation. The mutations responsible for benomyl resistance were determined in two spontaneous mutants, 8-18 and 8­19. One mutant 8-18 substituted glutamate for aspar­agine at position 33 and lysine for glutamine at position 134. The other mutant 8-19 showed alterations at three positions of $\beta$-tubulin arginine for tryptophan at position 21, lysine for asparagine at position 33, and phenylalanine for leucine at position 240. These data suggest that regions of $\beta$-tubulin containing amino acids 21, 33,134, and 240 interact to form the binding site of benomyl.

Taxonomy of $Ganoderma$ $lucidum$ from Korea Based on rDNA and Partial ${\beta}$-Tubulin Gene Sequence Analysis

  • Park, Young-Jin;Kwon, O-Chul;Son, Eun-Suk;Yoon, Dae-Eun;Han, Woo-Ri-Ja-Rang;Yoo, Young-Bok;Lee, Chang-Soo
    • Mycobiology
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    • v.40 no.1
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    • pp.71-75
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    • 2012
  • In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial ${\beta}$-tubulin gene sequence of the $Ganoderma$ species. The size of the ITS rDNA regions from different $Ganoderma$ species varied from 625 to 673 bp, and those of the partial ${\beta}$-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean $Ganoderma$ $lucidum$ strains belong in a single group along with a $G.$ $lucidum$ strain from Bangladesh.

Isolation and Sequencing of the cDNA Encoding ${\beta}-tubulin$ from Pleurotus sajor-caju (여름느타리버섯으로부터 ${\beta}-tubulin$ cDNA의 분리 및 염기서열 결정)

  • Kim, Beom-Gi;Shin, Pyung-Gyun;Jeong, Mi-Jeong;Park, Soo-Chul;Yoo, Young-Bok;Ryu, Jin-Chang;Kwon, Suk-Tae
    • The Korean Journal of Mycology
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    • v.25 no.1 s.80
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    • pp.1-5
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    • 1997
  • The cDNA encoding ${\beta}-tubulin$ of Pleurotus sajor-caju was isolated using an internal gene segment probe amplified by polymerase chain reaction (PCR) of genomic DNA and by cDNA library screening. The cDNA was consisted of 1560 nucleotides(nt), including a 5'-untranslation region (UTR) of 27nt, an open reading frame (ORF) of 1341nt, and a 3'-UTR of 191nt. The ORF encoded a protein of 446 amino acids(aa), which shows over 80% homology with ${\beta}-tubulins$ of other filamentous fungi. Southern hybridization analysis showed that there were two isotypes of ${\beta}-tubulin$ genes in P. sajor-caju. Through sequence analysis we found that ${\beta}-tubulin$ had a unusual $Cys^{165}$ residue, which might be a significant factor for the insensitivity of fungi to fungicide benomyl.

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First Report of Penicillium brasilianum and R. daleae Isolated from Soil in Korea

  • Cho, Hye-Sun;Hong, Seung-Beom;Go, Seung-Joo
    • Mycobiology
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    • v.33 no.2
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    • pp.113-117
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    • 2005
  • In this study, a total of 300 isolates of Penicillium and related teleomorphic genera were collected from soils of 17 locations in Korea from April to May, 2004. Ninety four isolates were identified as the species of Penicillium subgenus Furcatum based on cultural and morphological characteristics and ${\beta}-tubulin$ gene sequences. Among the specie's, Korean isolates of P. brasilianum Bat. and P. daleae K. M. Zalessky were phylogenetically identical to the reference species based on DNA sequence of the ${\beta}-tubulin$ gene. Here we described and illustrated P. brasilianum and P. daleae that are new in Korea.

Genotypic Characterization of Oak Wilt Pathogen Raffaelea quercus-mongolicae and R. quercivora Strains (참나무류에 시들음병을 일으키는 Raffaelea quercus-mongolicae와 R. quercivora의 유전적 특성)

  • Seo, Sang-Tae;Kim, Kyung-Hee;Lee, Sang-Hyun;Kwon, Yong-Nam;Shin, Chang-Hoon;Kim, Hye-Jeong;Lee, Sang-Yong
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.219-223
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    • 2010
  • Recently, the oak wilt diseases especially on Quercus mongolica, have been increasing in various districts of Korea. A collection of 38 strains of the oak wilt pathogen Raffaelea qeurcus-mongolicae and R. quecivora isolated from Quercus spp. in Korea and Japan was characterized by $\beta$-tubulin gene sequence and randomly amplified polymorphic DNA (RAPD) analysis. In cluster analysis based on $\beta$-tubulin gene sequence the strains were divided into 4 clusters, of which clusters 2 and 4 were composed of Japanese strains except for one Korean strain. RAPD analysis showed that they were also effectively differentiated by a strong RAPD fragments. On the basis of the two genetic analysis, significant differences were detected between Korean strains and Japanese strains.

The mRNA Expression of Radio-Sensitive Genes Exposed to Various Dosage of Ionizing Radiation in U-937 Cell (U-937 세포에서 이온화 방사선의 조사선량에 따른 감수성 유전자들의 발현 변화)

  • 김종수;임희영;오연경;김인규;강경선;윤병수
    • Toxicological Research
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    • v.20 no.1
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    • pp.21-29
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    • 2004
  • We used cDNA microarray to assess gene expression profiles in hematopoetic cell line, U-937, exposed to low doses of ionizing irradiation. The 1,000 DNA elements on this array were PCR-amplified cDNAs selected from named human cancer related genes. According to the strength of irradiation, the levels of some gene expression were increased or decreased as dose-dependent manner. The gene expressions of Tubulin alpha, protein kinase, interferon-alpha, -beta, -omega receptor and ras homolog gene family H were significantly increased. Especially, Tubulin gene was shown 2.5 fold up-regulated manner under stress of 500 rad irradiation than 200 rad. On the other hand, fibroblast growth factor 12 and four and a half LIM domains, etc. were significantly down-regu-lated. Also, tumor protein 53(TP53) related genes that p53 inducible protein, tumor protein 53-binding protein looks of little significance as radiation sensitive manner. The radio-sensitivity of tubulin gene etc. that we proposed could be useful to rapid and correct survey for the bio-damage by exposure to low dose irradiation.

Transformation of Metarhizium anisopliae by using pBRG-4 (pBRG-4를 이용한 Metarhizium anisopliae의 형질전환)

  • Lee, Dong-Gyu;Yeh, Wan-Hae;Hwang, Cher-Won;Kwon, Suk-Tae;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.41 no.3
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    • pp.219-223
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    • 1998
  • We have established a transformation system for entomopathogenic fungus, Metarhizium anisopliae, in order to develop mycoinsecticide by recombinant DNA techniques. Protoplasts of M. anisopliae would be transformed to a benomyl-resistant by introducing pBRG-4 plasmid DNA, which contains a ${\beta}-tubulin$ gene of Aspergillus flavus conferring resistance to benomyl and a pyr4 gene of Neurospora crassa, in the presence of 5% polyethylene glycol and 10 mM calcium chloride. Transformants occuring at a frequency of 10 colonies per $50\;{\mu}g$ pBRG-4 DNA grew on the $5\;{\mu}g/ml$ concentrations of benamyl, while the wild type was inhibited by $2.5\;{\mu}g/ml$. From the Southern analysis using genomic DNAs isolated from M. anisopliae transformants, the positive signals suggested that the ${\beta}-tubulin$ gene had integrated in the M. anisopliae genome by homologous recombination.

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A New Record of Penicillium pimiteouiense from Beach Soil in Malaysia

  • Teh, Li Yee;Latiffah, Zakaria
    • Mycobiology
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    • v.41 no.4
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    • pp.256-259
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    • 2013
  • Three isolates of Penicillium pimiteouiense were recovered from sandy beach soil samples in Penang Island, Peninsular Malaysia. All the isolates were identified based on morphological characteristics and phylogenetic analysis of internal transcribed spacer regions and ${\beta}$-tubulin gene. This is a first record of P. pimiteouiense in Malaysia.