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Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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Clinical Practice Guideline for Cardiac Rehabilitation in Korea

  • Kim, Chul;Sung, Jidong;Lee, Jong Hwa;Kim, Won-Seok;Lee, Goo Joo;Jee, Sungju;Jung, Il-Young;Rah, Ueon Woo;Kim, Byung Ok;Choi, Kyoung Hyo;Kwon, Bum Sun;Yoo, Seung Don;Bang, Heui Je;Shin, Hyung-Ik;Kim, Yong Wook;Jung, Heeyoune;Kim, Eung Ju;Lee, Jung Hwan;Jung, In Hyun;Jung, Jae-Seung;Lee, Jong-Young;Han, Jae-Young;Han, Eun Young;Won, Yu Hui;Han, Woosik;Baek, Sora;Joa, Kyung-Lim;Lee, Sook Joung;Kim, Ae Ryoung;Lee, So Young;Kim, Jihee;Choi, Hee Eun;Lee, Byeong-Ju;Kim, Soon
    • Journal of Chest Surgery
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    • v.52 no.4
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    • pp.248-329
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    • 2019
  • Background: Though clinical practice guidelines (CPGs) for cardiac rehabilitation (CR) are an effective and widely used treatment method worldwide, they are as yet not widely accepted in Korea. Given that cardiovascular disease is the second leading cause of death in Korea, it is urgent that CR programs be developed. In 2008, the Government of Korea implemented CR programs at 11 university hospitals as part of its Regional Cardio-Cerebrovascular Center Project, and 3 additional medical facilities will be added in 2019. In addition, owing to the promotion of CR nationwide and the introduction of CR insurance benefits, 40 medical institutions nationwide have begun CR programs even as a growing number of medical institutions are preparing to offer CR. The purpose of this research was to develop evidence-based CPGs to support CR implementation in Korea. Methods: This study is based on an analysis of CPGs elsewhere in the world, an extensive literature search, a systematic analysis of multiple randomized control trials, and a CPG management, development, and assessment committee comprised of 33 authors-primarily rehabilitation specialists, cardiologists, and thoracic surgeons in 21 university hospitals and 2 general hospitals. Twelve consultants, primarily rehabilitation, sports medicine, and preventive medicine specialists, CPG experts, nurses, physical therapists, clinical nutritionists, and library and information experts participated in the research and development of these CPGs. After the draft guidelines were developed, 3 rounds of public hearings were held with staff members from relevant academic societies and stakeholders, after which the guidelines were further reviewed and modified. Results: CR involves a more cost-effective use of healthcare resources relative to that of general treatments, and the exercise component of CR lowers cardiovascular mortality and readmission rates, regardless of the type of coronary heart disease and type and setting of CR. Conclusion: Individualized CR programs should be considered together with various factors, including differences in heart function and lifestyle, and doing so will boost participation and adherence with the CR program, ultimately meeting the final goals of the program, namely reducing the recurrence of myocardial infarction and mortality rates.

Construction of Genetic Linkage Map and Identification of Quantitative Trait Loci in Populus davidiana using Genotyping-by-sequencing (Genotyping-by-sequencing 기법을 이용한 사시나무(Populus davidiana) 유전연관지도 작성 및 양적형질 유전자좌 탐색)

  • Suvi Kim;Yang-gil Kim;Dayoung Lee;Hye-jin Lee;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.1
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    • pp.40-56
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    • 2023
  • Tree species within the Populus genus grow rapidly and have an excellent capacity to absorb carbon, conferring substantial ability to effective purify the environment. Poplar breeding can be achieved rapidly and efficiently if a genetic linkage map is constructed and quantitative trait loci (QTLs) are identified. Here, a high-density genetic linkage map was constructed for the control pollinated progeny using the genotyping-by-sequencing (GBS) technique, which is a next-generation sequencing method. A search was also performed for the genes associated with quantitative traits located in the genetic linkage map by examining the variables of height and diameter at root collar, and resilience to insect damage. The height and diameter at root collar were measured directly, while the ability to recover from insect damage was scored in a 4-year-old breeding population of aspen hybrids (Odae19 × Bonghyeon4 F1) established in the research forest of Seoul National University. After DNA extraction, paternity was confirmed using five microsatellite markers, and only the individuals for which paternity was confirmed were used for the analysis. The DNA was cut using restriction enzymes and the obtained DNA fragments were prepared using a GBS library and sequenced. The analyzed results were sorted using Populus trichocarpa as a reference genome. Overall, 58,040 aligned single-nucleotide polymorphism (SNP) markers were identified, 17,755 of which were used for mapping genetic linkages. The genetic linkage map was divided into 19 linkage groups, with a total length of 2,129.54 cM. The analysis failed to identify any growth-related QTLs, but a gene assumed to be related to recovery from insect damage was identified on linkage group (chromosome) 4 through genome-wide association study.

Assessment of Physiological Activity of Entomopathogenic Fungi with Insecticidal Activity Against Locusts (풀무치에 대하여 살충활성을 보유한 곤충병원성 진균의 생리활성 평가)

  • Lee, Mi Rong;Kim, Jong Cheol;Lee, Se Jin;Kim, Sihyeon;Lee, Seok Ju;Park, So Eun;Lee, Wang Hyu;Kim, Jae Su
    • Korean journal of applied entomology
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    • v.56 no.3
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    • pp.301-308
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    • 2017
  • Locusts, Locusta migratoria (Orthoptera: Acrididae) are periodical unpredictable agricultural pests worldwide and cause serious damage to crop production; however, little consideration has been given to the management of this pest. Herein, we constructed a locust-pathogenic fungal library and confirmed that some fungi could be used as resources for locust management. First, the entomopathogenic fungi were collected from sampled soils using a Tenebrio molitor-based baiting system. For the locust assay, a locust colony was obtained from the National Institute of Agricultural Science and Technology. A total of 34 entomopathogenic fungal granules, which were produced by solid cultures, were placed in the plastic insect-rearing boxes (2 g/box) and nymphs of locust were contained in the box. In 3-7 days, mycosis was observed on the membranous cuticles of the head, abdomen, and legs of locusts. In particular, Metarhizium anisopliae, M. lepidiotae, and Clonostachys rogersoniana exhibited high virulence against the locust. Given that the 34 isolates could be used in field applications, their conidial production and stability (thermotolerance) were further characterized. In the thermotolerance assay, Paecilomyces and Purpureocillium isolates had higher thermotolerance than the other isolates. Most of the fungal isolates produced ca. >$1{\times}10^8conidia/g$ on millet grain medium. In a greenhouse trial, the granular application of M. anisopliae isolate on the soil surface resulted in 85.7% control efficacy. This work suggests that entomopathogenic fungi in a granular form can be effectively used to control the migratory locust.

Analysis of Research Trends Related to Christian Picture Books : Focusing on Domestic Dissertations (기독교 그림책 관련 연구 동향 분석 : 국내 학위 논문 중심으로)

  • Kim, Minjung
    • Journal of Christian Education in Korea
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    • v.68
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    • pp.245-277
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    • 2021
  • The purpose of this study was to investigate the trend of Christian picture book-related research. The purpose of this study is to present basic data for various and balanced research and development in the Christian picture book field by analyzing the research period, research content, and research method related to Christian picture books. For this study, 45 domestic master's and doctoral dissertations were extracted through the National Assembly Library and the Academic Research Information Service (RISS) with the keywords of 'Christian picture book', 'Bible picture book', 'Christian story', and 'Bible story'. The frequency and percentage were calculated by analyzing Christian picture book-related studies according to four criteria: research period, research content, research method, and research subject. As a result of the study, first, the trend of Christian picture book research papers by research period from 1999 to 2021 was 43 master's articles (95.6%) and 2 doctoral articles (4.4%), focusing on Christian picture book-related studies. Second, the trend by research content was found to be 12 basic studies (26.6%) and 33 practical studies (73.4%). Research related to Christian picture books is being actively conducted focusing on practical research rather than basic research. Third, the trend by research method was in the order of 33 quantitative studies (73.4%), 11 literature studies (24.4%), and 1 qualitative study (2.2%). Research related to Christian picture books is centered on quantitative research, and literature research and qualitative research account for a relatively low proportion. Fourth, as for the trends by study subject, there were 35 human subjects (77.8%) and 10 physical subjects (22.2%). Among human subjects, 33 single subjects (73.4%) and 2 mixed subjects (4.4%) were found, and among single subjects, 30 studies (66.7%) targeting children were high. In other words, research on Christian picture books had a higher proportion of studies with children as a single subject than mixed subjects between children and children, children and teachers, and between children and parents.

Blended IT/STEM Education for Students in Developing Countries: Experiences in Tanzania (개발도상국 학생들을 위한 블랜디드 IT/STEM교육: 탄자니아에서의 경험 및 시사점)

  • Yoon Rhee, Ji-Young;Ayo, Heriel;Rhee, Herb S.
    • Journal of Appropriate Technology
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    • v.6 no.2
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    • pp.151-162
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    • 2020
  • Education is one of the priority sectors specified in Tanzania, and it has committed to provide 11 years of compulsory free basic education for all from pre-primary to lower secondary level. Despite the Government's efforts to provide free basic education to all children, there are 2.0 million (23.2 per cent) out of 8.5 million children at the primary school age of 7-13, who are out of school in Tanzania. The ICT class should be offered as a regular class in all secondary schools in Tanzania, recommended by the ministry of education. However, many schools are struggling to implement this mandate. Most of schools offer the ICT class with theory without any real hardware. Some schools were given with computers but they were not maintained for operation. There is a huge task to make ICT education universal. Main issues include: remoteness (off-grid area), lack of ICT teachers, lack of resources such as hardware, infrastructure, and lack of practical lessons or projects to be used at schools. An innovative blended ICT/STEM education program is being conducted not only for Tanzanian public and private/international schools, but also for out-of-school adolescents through institutions, NGO centers, home visits and at the E3 Empower academy center. For effective STEM education to take place and remain sustainable, more practical curriculum, and close-up teacher support need to be accompanied concurrently. Practical, project-based simple coding lessons have been developed and employed that students experience true learning. The effectiveness of the curriculum has been demonstrated in various project centers, and it showed that students are showing new interests in exploring new discovery, even though this was a totally new area for them. It has been designed for an easy replication, thus students who learned can repeat the lessons themselves to other students. The ultimate purpose of this project is to have IT education offered as universally as possible throughout the whole Tanzania. Quality education for all children is a key for better future for all. Previously it was hoped that education with discipline will improve the active learning. But now more than ever, we believe that children have the ability to learn on their own with given proper STEM education tools, guidelines and environment. This gives promising hope to all of us, including those in the developing countries.

Molecular Cloning of Novel Genes Specifically Expressed in Snailfish, Liparis tanakae (꼼치, Liparis tanakae에서 특이하게 발현되는 새로운 유전인자의 검색)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.4 no.1
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    • pp.67-77
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    • 2000
  • Snailfish usually lives at the bottom of the sea and showed typical retrogressive change with specialized tissue structures of skin and skeletons. In order to obtain the specific genes of snailfish, highly expressed in the body, we made subtracted cDNA library and analyzed 200 clones. Totally 200 clones were obtained and sequenced, and among them 62 clones were turned out to be homologous to the known gene, i.e., thioesterase (9), myosin (8), creatine kinase (7), skeletal alpha-actin (6), parvalbumin b (5), ribosomal protein (5), type I collagen (3), muscle troponin (3), dopamine receptor (2), histatin (2), and heat shock protein (2), cystatin (1), lectin (1), statherin (1), secretory carrier membrane protein (1), keratin type I (1), desmin (1), chloroplast (1), muscle tropomyosin (1), reticulum calcium ATPase (1), ribonucleoprotein (1). The remaining 138 clones were low homologous or non-redundant genes through Genbank search. Especially 5 clones were novel and specifically expressed in the body tissues of Snailfish by in situ hybridization. Therefore, we analysed these 5 clones to identify the C-terminal protein structures and motifs, and partly defined the roles of these proteins in comparison with the expression patterns by in situ hybridization. C9O-77, about 5000 bp, was supposed to be a matrix protein expressed strongly positive in epithelium, myxoid tissue, fibrous tissue and collagenous tissue. C9O-116, about 1500 bp, was supposed to be a transmembrane protein which was weakly expressed in the fibrous tissue, epithelium tissue, and myxoid tissue, but strong in muscle tissue. C9O-130, about 1200 bp, was supposed to be an intracytoplasmic molecule usually in the epithelial cells. C9O-161, about 2000 bp, was weakly expressed in epithelium, muscle tissue and myxoid tissue, but specially strong in epithelium. C9O-171, about 1000 bp, was supposed to be a transcription factor containing zinc finger like domain, which was intensely expressed in the epithelium, muscle tissue, fibrous tissue, and in collagenous tissue.

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Characterization of the cloned RNA1 gene of Saccharomyces cerevisiae (Cloning된 효모의 RNAI 유전자의 특성에 관하여)

  • Song, Young-Hwan;Kim, Dae-Young;Kim, Jin-Kyung
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.93-101
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    • 1993
  • The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

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Heparanase mRNA and Protein Expression Correlates with Clinicopathologic Features of Gastric Cancer Patients: a Meta-analysis

  • Li, Hai-Long;Gu, Jing;Wu, Jian-Jun;Ma, Chun-Lin;Yang, Ya-Li;Wang, Hu-Ping;Wang, Jing;Wang, Yong;Chen, Che;Wu, Hong-Yan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8653-8658
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    • 2016
  • Background: Heparanase is believed to be involved in gastric carcinogenesis. However, the clinicopathologic features of gastric cancer with high heparanase expression remain unclear. Aim : The purpose of this study was to comprehensively and quantitatively summarize available evidence for the use of heparanase mRNA and protein expression to evaluate the clinicopathological associations in gastric cancer in Asian patients by meta-analysis. Materials and Methods: Relevant articles listed in MEDLINE, CNKI and the Cochrane Library databases up to MARCH 2015 were searched by use of several keywords in electronic databases. A meta-analysis was performed to clarify the impact of heparanase mRNA and protein on clinicopathological parameters in gastric cancer. Combined ORs with 95%CIs were calculated by Revman 5.0, and publication bias testing was performed by stata12.0. Results: A total of 27 studies which included 3,891 gastric cancer patients were combined in the final analysis. When stratifying the studies by the pathological variables of heparanase mRNA expression, the depth of invasion (633 patients) (OR=4.96; 95% CI=2.38-1.37; P<0.0001), lymph node metastasis (639 patients) (OR=6.22; 95%CI=2.70-14.34, P<0.0001), and lymph node metastasis (383 patients) (OR=6.85; 95% CI=2.04-23.04; P=0.002) were all significant. When stratifying the studies by the pathological variables of heparanase protein expression, this was the case for depth of invasion (1250 patients) (OR=2.76; 95% CI=1.52-5.03; P=0.0009), lymph node metastasis (1178 patients) (OR=4.79 ; 95% CI=3.37-6.80, P<0.00001), tumor size (727 patients) (OR=2.06 ; 95% CI=1.31-3.23; P=0.002) (OR=2.61; 95% CI=2.09-3.27; P=0.000), and TNM stage (1233 patients) (OR=6.85; 95% CI=2.04-23.04; P=0.002). Egger's tests suggested publication bias for depth of invasion, lymph node metastasis, lymph node metastasis and tumor size of heparanase mRNA and protein expression. Conclusions: This meta-analysis suggests that higher heparanase expression in gastric cancer is associated with clinicopathologic features of depth of invasion, lymph node metastasis and TNM stage at mRNA and protein levels, and of tumor size only at the protein level. Egger's tests suggested publication bias for these clinicopathologic features of heparanase mRNA and protein expression, and which may be caused by shortage of relevant studies. As a result, although abundant reports showed heparanase may be associated with clinicopathologic features in gastric cancer, this meta-analysis indicates that more strict studies were needed to evaluate its clinicopathologic significance.

Development of System-Wide Functional Analysis Platform for Pathogenicity Genes in Magnaporthe oryzae

  • Park, Sook-Young;Choi, Jaehyuk;Choi, Jaeyoung;Kim, Seongbeom;Jeon, Jongbum;Kwon, Seomun;Lee, Dayoung;Huh, Aram;Shin, Miho;Jung, Kyungyoung;Jeon, Junhyun;Kang, Chang Hyun;Kang, Seogchan;Lee, Yong-Hwan
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.9-9
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    • 2014
  • Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.

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