• Title/Summary/Keyword: over-expression vector

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Isolation and Characterization of Zymomonas mobilis DNA Fragments Showing Promoter Activity in Escherichia coli (Escherichia coli에서 Promoter 활성을 보이는 Zymomonas mobilis DNA 조각의 분리와 분석)

  • Kim, Eun-Joon;Yoon, Ki-Hong;M.Y. Pack
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.600-605
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    • 1989
  • For the purpose of isolation of the Zymomonas mobilis DNA fragments showing promoter activity in Escherichia coli, a promoter screening vector, PCMT215 was constructed by transferring a promoterless chloramphenicol acetyltransferase (CAT) gene of pYEJ001 into pMT21 which contains $\beta$-lactamase gene and multiple cloning sites. A library of Z, mobilis Sau3AI DNA fragments was constructed in E. coli using the newly constructed pCMT215. Fourteen clones showing resistance to chloramphenicol ranging in concentration from 30 to 750 $\mu$g/$m\ell$ were selected. From five clones of them, the Z. mobilis DNA fragments expressing CAT gene of the recombinant plasmids were sequenced and then sites of transcriptional initiation were identified. The nucleotide sequences of the cloned DNA shared AT rich regions, poly A's or T's stretches and palindromic regions. The positions of transcriptional initiation for CAT gene occurred at more than one site spaced over by 4 to 190 base pairs on the cloned fragments in E. coli.

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The characterization of transgenic Chrysanthemum under low temperature condition (저온저항성 유전자가 도입된 국화 형질전환체 특성)

  • Choi, In-Young;Han, Soo-Gon;Kang, Chan-Ho;Song, Young-Ju;Lee, Wang-Hyu
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.55-61
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    • 2008
  • Previous studies on genetic transformation of chrysanthemum using cold regulated gene (BN115) have been conducted and the PCR and Real-Time PCR based method to determine the presence of the transferred cold regulated gene in the chrysanthemum was established. To check whether over-expression of BN115 gene in transgenic chrysanthemum will enhance their tolerance to cold stress, the transgenic chrysanthemum were grown under low temperature condition and several cold signalling including growth characteristics, stoma size and shape, SPAD value and ion leakage test were investigated. The transgenic chrysanthemum in the low temperature growth chamber grow much faster in term of the height, number and size of the leaves than those of wild-type plants and damage of transgenic plant caused by the low temperature was much less than that of wild-type plants. The stoma type and size of transgenic plant leaves grown at $5^{\circ}C$ were much similar to of wild-type plant cultured on $25^{\circ}C$ It has been found that SPAD value of transgenic plants was much higher than those of wild-type, but the EC density being lower under low temperature condition.

Characterization of Transgenic Tall Fescue Plants Expressing Two Antioxidant Genes in Response to Environmental Stresses (두 가지 항산화유전자를 동시에 발현시킨 형질전환 톨 페스큐 식물체의 환경스트레스에 대한 내성 특성 해명)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Seo, Sung;Kwak, Sang-Soo;Kwon, Suk-Yoon;Yun, Dae-Jin;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.2
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    • pp.109-116
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    • 2007
  • Environmental stress is the major limiting factor in plant productivity. As an effort to solve the global food and environmental problems using the plant biotechnology, we have developed transgenic tall fescue (Festuca arundinacea Schreb.) plants via Agrobacterium-mediated gene transfer method. To develop transgenic tall fescue plants with enhanced tolerance to the environmental stresses, both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) genes were incorporated in a pIG121 binary vector and the both of the genes were controlled separately by an oxidative stress-inducible sweet potato peroxidase 2 (SWPA2) premoter expressed in chloroplasts. Leaf discs of transgenic plants showed 10-30% less damage compared to the wild-type when they exposed to a wide range of environmental stresses including methyl viologen (MV), $H_2O_2$ and heavy metals. In addition, when $200{\mu}M$ MV was sprayed onto the whole plants, transgenic plants showed a significant reduction of visible damage compared to wild-type plants that were almost damaged. These results suggest that over expression of CuZnSOD and APX genes in transgenic plants might be a useful strategy to protect the crops against a wide range of environmental stresses.

Study on Expression and Characterization of HRD3 Gene Related DNA Repair from Eukaryotic Cells (진핵세포에서 DNA 회복에 관련된 HRD3 유전자의 분리, 발현 및 특성 연구)

  • Shin, Su-Hwa;Park, In-Soon
    • Journal of Life Science
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    • v.14 no.2
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    • pp.325-330
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    • 2004
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA and DNA-RNA helicase activities. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (Homologue of RAD3 gene). The over-expressed HRD3 protein was estimated to be a 75 kDa in size which is in good agreement with the estimated by the nucleotide sequence of the cloned gene. Two-dimensional gel electrophoresis showed that a number of other protein spots dramatically disappeared when the HRD3 protein was overexpressed. The overexpressed RAD3 protein showed a toxicity in E. coli host, suggesting that this protein may be involved in the inhibition of protein synthesis and/or degradation of host protein. To determine which part of HRD3 gene contributes to the toxicity in E. coli, various fusion plasmids containing a partial sequence of HRD3 and lac'Z gene were constructed. These results suggest that the C-terminal domain of HRD3 protein may be important for both toxic effect in E. coli and for its role in DNA repair in S. pombe.

Selective Word Embedding for Sentence Classification by Considering Information Gain and Word Similarity (문장 분류를 위한 정보 이득 및 유사도에 따른 단어 제거와 선택적 단어 임베딩 방안)

  • Lee, Min Seok;Yang, Seok Woo;Lee, Hong Joo
    • Journal of Intelligence and Information Systems
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    • v.25 no.4
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    • pp.105-122
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    • 2019
  • Dimensionality reduction is one of the methods to handle big data in text mining. For dimensionality reduction, we should consider the density of data, which has a significant influence on the performance of sentence classification. It requires lots of computations for data of higher dimensions. Eventually, it can cause lots of computational cost and overfitting in the model. Thus, the dimension reduction process is necessary to improve the performance of the model. Diverse methods have been proposed from only lessening the noise of data like misspelling or informal text to including semantic and syntactic information. On top of it, the expression and selection of the text features have impacts on the performance of the classifier for sentence classification, which is one of the fields of Natural Language Processing. The common goal of dimension reduction is to find latent space that is representative of raw data from observation space. Existing methods utilize various algorithms for dimensionality reduction, such as feature extraction and feature selection. In addition to these algorithms, word embeddings, learning low-dimensional vector space representations of words, that can capture semantic and syntactic information from data are also utilized. For improving performance, recent studies have suggested methods that the word dictionary is modified according to the positive and negative score of pre-defined words. The basic idea of this study is that similar words have similar vector representations. Once the feature selection algorithm selects the words that are not important, we thought the words that are similar to the selected words also have no impacts on sentence classification. This study proposes two ways to achieve more accurate classification that conduct selective word elimination under specific regulations and construct word embedding based on Word2Vec embedding. To select words having low importance from the text, we use information gain algorithm to measure the importance and cosine similarity to search for similar words. First, we eliminate words that have comparatively low information gain values from the raw text and form word embedding. Second, we select words additionally that are similar to the words that have a low level of information gain values and make word embedding. In the end, these filtered text and word embedding apply to the deep learning models; Convolutional Neural Network and Attention-Based Bidirectional LSTM. This study uses customer reviews on Kindle in Amazon.com, IMDB, and Yelp as datasets, and classify each data using the deep learning models. The reviews got more than five helpful votes, and the ratio of helpful votes was over 70% classified as helpful reviews. Also, Yelp only shows the number of helpful votes. We extracted 100,000 reviews which got more than five helpful votes using a random sampling method among 750,000 reviews. The minimal preprocessing was executed to each dataset, such as removing numbers and special characters from text data. To evaluate the proposed methods, we compared the performances of Word2Vec and GloVe word embeddings, which used all the words. We showed that one of the proposed methods is better than the embeddings with all the words. By removing unimportant words, we can get better performance. However, if we removed too many words, it showed that the performance was lowered. For future research, it is required to consider diverse ways of preprocessing and the in-depth analysis for the co-occurrence of words to measure similarity values among words. Also, we only applied the proposed method with Word2Vec. Other embedding methods such as GloVe, fastText, ELMo can be applied with the proposed methods, and it is possible to identify the possible combinations between word embedding methods and elimination methods.