• Title/Summary/Keyword: Dominant selectable marker

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The Use of Aureobasidin A Resistant Gene as the Dominant Selectable Marker for the Selection of Industrial Yeast Hybrid (산업용 효모 Hybrid의 선별을 위한 우성선별표지로서의 Aureobasidin A 내성유전자의 이용)

  • Jeon, Han-Taek;Park, Uhn-Mee;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.111-118
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    • 2011
  • For the strain improvement of the industrial polyploid yeast strain through hybridization and protoplast fusion, a dominant selection marker other than a recessive marker such as the auxotrophic marker was required for the selection of the resulting hybrids. In the present investigation, the aureobasidin A resistant gene was tested in relation to whether it can be used as the dominant selectable marker for the isolation of hybrids of the yeast Saccharomyces. The plasmid pAUR112, carrying the gene responsible for resistance to aureobasidin A, was introduced into the haploid yeast strain K114/YIp. From the rare-mating between polyploid C6 and haploid K114/YIp carrying pAUR112, many hybrids were obtained from the agar medium containing 0.5 ${\mu}g$/ml of aureobasidin A. The hybrids exhibited characteristics derived from both of the parental strains; and the cell sizes of the hybrids were larger than those of the parental strains. These results showed that the aureobasidin A resistant gene could be successfully used as the dominant selectable marker for the isolation of yeast hybrids resulting from rare-mating.

Expression of Schwanniomyces occidentalis $\alpha-Amylase$ Gene in Saccharomyces cerevisiae var. diastaticus

  • Park, Jeong-Nam;Shin, Dong-Jun;Kim, Hee-Ok;Kim, Dong-Ho;Lee, Hwang-Hee;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.668-671
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    • 1999
  • The gene encoding Schwanniomyces occidentalis $\alpha-amylase$(AMY) was introduced into Saccharomyces cerevisiae var. diastaticus which secreted only glucoamylase, by using a linearized yeast integrating vector to develop stable strains with a capability of secreting $\alpha-amylase$and glucoamylase simultaneously. A dominant selectable marker, the geneticin(G418) resistance gene (Gt^r$), was cloned into a vector to screen wild-type diploid transformants harboring the AMY gene. The amylolytic activities of transformants were about 3-7 times higher than those of the recipient strains. When grown in nonselective media, the transformants with the linearized integrating vector containing the AMY gene exhibited almost all of the mitotic stability after 100 generations.

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Transformation of Coprinus congregatus with a Linearized Plasmid Vector to Phosphinothricin Resistance (Coprinus congregatus에서 선형으로 전환한 plasmid DNA를 사용하여 phosphinothricin 저항성에 대한 형질전환)

  • Leem, Young-Eun;Kim, Soon-ja;Choi, Hyoung-Tae
    • Korean Journal of Microbiology
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    • v.33 no.4
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    • pp.274-276
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    • 1997
  • Transformation of Coprinus congregatus with a linearized plasmid has been successfully carried out using phosphinothricin resistance gene as a dominant selectable marker. The transforming frequency was about 500 transformants per microgram of DNA using the protoplast-$CaCl_2$ method. The transforming vector pBARGEM 7-1 which had the phosphinothricin resistance gene was detected in the restriction enzyme fragments of chromosomal DNA from a transformant by Southern hybridization.

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Transfection and Expression of Reconstructed Genes within Baculoviral Vectors (Baculovirus 벡터내 재구성된 유전자의 전이와 발현)

  • Sa, Young-Hee;Choi, hang-Shik;Lee, Ki Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.05a
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    • pp.588-591
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    • 2018
  • Baculovirus was originally isolated from the alfalfa looper and contains a 134-kbp genome with 154 open reading frames (ORF). The major capsid protein VP39 together with some minor proteins forms the nucleocapsid ($21nm{\times}260nm$) that encloses the DNA with p6.9 protein. They are double-stranded, circular, supercoiled DNA molecules in a rod-shaped capsid. Wild-type baculoviruses exhibit both lytic and occluded life cycles that develop independently throughout the three phases of virus replication. Recombinant baculoviruses can transfer their vectors and express their recombinant proteins in a wide range of mammalian cell types. Especially, inclusion of a dominant selectable marker in these baculoviral vectors can express diverse recombinant genes in many cells. Baculoviral vectors were reconstructed with cytomegalovirus (CMV) promoter,uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) gene and so on. These reconstructed vectors were infected into various cell and cell lines. We performed transfection and expression of these recombinant vectors comparison with other control vectors. From this study, we knew that transfection and expression of these recombinant vectors have higher efficacy than any control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

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Transgenic Tomato Plants That Overexpress Superoxide Dismutase in Fruits (토마토 과실에서 Superoxide Dismutase를 고발현하는 형질전환 식물체)

  • Park, Eun-Jeong;Lee, Haeng-Soon;Kwon, Suk-Yoon;Choi, Kwan-Sam;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.7-13
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    • 2002
  • Superoxide dismutase (SOD) plays an important role in cellular defense against oxidative stress in plants. We have developed transgenic tomato plants overexpressing a cassava SOD in fruits. Three transgenic tomato plants (one from cv. Pink forcer and two from cv. Koko) using a new vector system, ASOp :: . mSOD1/pBI101, harboring ascorbate oxidase promoter (ASOp) expressing dominantly in cucumber fruits, CuZnSOD cDNA (mSOD1) isolated from cultured cells of cassava, and nptll gene as a selectable marker were successfully developed. SOD specific activity (units/mg protein) in transgenic fruits of both cultivars was increased with maturation of the fruits. SOD specific activity of well-mature fruits in transgenic Pink forcer and Koko showed approximately 1.6 and 2.2 times higher than control fruits, respectively. The strength of SOD isoenzyme bands well reflected the SOD activity during the fruit maturation. These results suggested that SOD gene was properly introduced into tomato fruits in a fruit-dominant expression manner by ASO promoter.