• Title/Summary/Keyword: That-Complementation

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cis-Prenyltransferase interacts with a Nogo-B receptor homolog for dolichol biosynthesis in Panax ginseng Meyer

  • Nguyen, Ngoc Quy;Lee, Sang-Choon;Yang, Tae-Jin;Lee, Ok Ran
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.403-410
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    • 2017
  • Background: Prenyltransferases catalyze the sequential addition of isopentenyl diphosphate units to allylic prenyl diphosphate acceptors and are classified as either trans-prenyltransferases (TPTs) or cis-prenyltransferases (CPTs). The functions of CPTs have been well characterized in bacteria, yeast, and mammals compared to plants. The characterization of CPTs also has been less studied than TPTs. In the present study, molecular cloning and functional characterization of a CPT from a medicinal plant, Panax ginseng Mayer were addressed. Methods: Gene expression patterns of PgCPT1 were analyzed by quantitative reverse transcription polymerase chain reaction. In planta transformation was generated by floral dipping using Agrobacterium tumefaciens. Yeast transformation was performed by lithium acetate and heat-shock for $rer2{\Delta}$ complementation and yeast-two-hybrid assay. Results: The ginseng genome contains at least one family of three putative CPT genes. PgCPT1 is expressed in all organs, but more predominantly in the leaves. Overexpression of PgCPT1 did not show any plant growth defect, and its protein can complement yeast mutant $rer2{\Delta}$ via possible protein-protein interaction with PgCPTL2. Conclusion: Partial complementation of the yeast dolichol biosynthesis mutant $rer2{\Delta}$ suggested that PgCPT1 is involved in dolichol biosynthesis. Direct protein interaction between PgCPT1 and a human Nogo-B receptor homolog suggests that PgCPT1 requires an accessory component for proper function.

Molecular Cloning and Characterization of Outer Envelope Membrane Protein from Salicornia herbacea (퉁퉁마디로부터 색소체 외막 단백질 유전자의 분리 및 발현분석)

  • Ermawati Netty;Cha, Joon-Yung;Liang, Yingshi;Jung, Min-Hee;Shin, Dongjin;Lee, Byung-Hyun;Lee, Kon-Ho;Son, Daeyoung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.273-278
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    • 2004
  • Complementary DNA encoding chloroplast outer envelope membrane protein (OEP) from the halophyte Salicornia herbacea has been cloned and sequenced. The full length cDNA is 596 bp and encodes a polypeptide of 91 amino acid residues with a molecular mass of 8.9 kDa. The expression level of ShOEP increased by salt, drought and ABA treatments. ShOEP expression was largely induced in roots and shoots by high salts. The biological function of ShOEP was examined by yeast complementation. ShOEP can suppress Na$^{+}$ sensitivity of yeast mutant (cnb$\Delta$) in the presence of salt. These results suggest that ShOEP is a salt inducible gene and may have functions in the regulation of plant salt stress.ant salt stress.

Cloning of Acetyl CoA Carboxylase (fabE) in Escherichia coli (대장균의 acetyl CoA carboxylase유전자의 클로닝)

  • Park, Wan;Song, Bang-Ho;Hong, Soon-Duk
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.181-186
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    • 1986
  • A defective lambda transducing phase carrying acetyl CoA carboxylase gene (fabE) from Escherichia coli chromosome (72 min on the current linkage map) has been isolated. A restriction map of the chromosomal region from defective transducing phage was established by digestion with combination of the restriction enzymes. No cleavage site for the enzyme EcoRI was found in this region. Restriction fragments were cloned from defective transducing phage into high copy number plasmid vector pACYC184 to generate hybrid plasmids which were capable of complementation of fabE temperature sensitive mutation. We show here that the fabE gene is located on a 3.4 megadalton Bam HI-SalI fragment with a HindIII site, which lies within the 7.4 megadalton BglIIfragment, by complementation analysis.

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Understanding the Soteriology of the Film : Focusing on the 'The Human Complementation Project' (의 구원관 연구: '인류보완계획'을 중심으로)

  • Lee, Kil-Yong
    • Cartoon and Animation Studies
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    • s.19
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    • pp.1-16
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    • 2010
  • The purpose of this Paper is a studying the soteriology of the film 'The End of Evangelion'(1997). In many religions the human being is recognized as negative and defect existence, on this count human being needs the salvation. Therefor the word salvation implies the meaning of liberation or overcoming. On this point Christianity, Buddhism, and Islam are of one accord. The christianity sees the original sin as the human limits, and the buddhism points out the ignorance(無明, ajnana) of human being. The soteriology of the film 'The End of Evangelion' is similar to the above-mentioned religions. For that the terminology of 'the Human Complementation Project' is used in film. In many religions the salvation is fulfilled in this world, in other words 'now' and 'here' are emphasized fot the accomplishment of the salvation. In this point the film 'the End of Evangelion' stands on the same road with the other religions.

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Meta-analysis of Excision Repair Cross-complementation Group 1 (ERCC1) Association with Response to Platinum-based Chemotherapy in Ovarian Cancer

  • Li, Feng-Ying;Ren, Xiao-Bin;Xie, Xin-You;Zhang, Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7203-7206
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    • 2013
  • Recent studies suggested that the ovarian cancers with negative excision repair cross-complementation group 1 enzyme (ERCC1) expression have a better response to platinum-based chemotherapy than those with positive ERCC1 expression. The objective of this study was to evaluate whether ERCC1 expression is associated with response to platinum-based chemotherapy in ovarian cancers. MEDLINE, PubMed, Web of Science and CNKI databases were used for searching studies relating to ERCC1 protein expression and response to platinum-based chemotherapy in ovarian cancers. Statistical analysis was based on the method for a fixed effects meta-analysis. Pooled odds ratios (ORs) with 95% confidence intervals for ERCC1 protein expression and response to platinum-based chemotherapy were generated. Publication bias was investigated with Begg's test. Five studies involving 306 patients with ovarian cancer were included. Compared to patients with positive ERCC1 expression, those with negative ERCC1 expression had a better response to platinum-based chemotherapy. The pooled OR was 5.264 (95% CI: 2.928-9.464, P < 0.001) and publication bias was not found (P = 0.904). The result was similar in both in Asians and Caucasians (P < 0.001 and P = 0.028, respectively). ERCC1 protein expression status is significantly associated with response to platinum-based chemotherapy in ovarian cancers.

Generation of Embryonic Stem Cell-derived Transgenic Mice by Using Tetraploid Complementation

  • Park, S.M.;Song, S.J.;Uhm, S.J.;Cho, S.G.;Park, S.P.;Lim, J.H.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1641-1646
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    • 2004
  • The objective of this study was to generate transgenic mice expressing human resistin gene by using the tetraploidembryonic stem (ES) cell complementation method. Human resistin gene was amplified from human fetal liver cDNA library by PCR, cloned into $pCR^{(R)}$ 2.1 $TOPO^{(R)}$ vector and constructed in pCMV-Tag4C vector. Mammalian expression plasmid containing human resistin was transfected into D3-GL ES cells by Lipofectamine 2,000, and then after 10-12 days of transfection, the human resistin-expressing cells were selected with G418. In order to produce tetraploid embryos, blastomeres of diploid embryos at the two-cell stage were fused with two times of electric pulse using 60 V 30 $\mu$sec (fusion rate: 2,114/2,256, 93.5%) and cultured up to the blastocyst stage (development rate: 1,862/2,114, 94.6%). The selected 15-20 ES cells were injected into tetraploid blastocysts, and then transferred into the uteri of E 2.5 d pseudopregnant recipient mice. To investigate the gestation progress, two E 19.5 mused fetuses were recovered by Cesarean section of which one fetus was confirmed to contain human resistin gene by genomic DNA-PCR. Therefore, our findings demonstrate that tetraploid-ES mouse technology can be considered as a useful tool to produce transgenic mice for the rapid analysis of gene function in vivo.

Synthetic Lethal Mutations with spmex67 of Schizosaccharomyces pombe in the Mediation of mRNA Export

  • Yoon, Jin-Ho
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.115-120
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    • 2003
  • Mex67p/Tap are evolutionally conserved mRNA export factors. To identify mutations in genes that are functionally linked to mex67 with respect to mRNA export, we used a synthetic lethal genetic screen in Schizosaccharomyces pombe. Three synthetic lethal mutants were isolated and mutations in these mutants defined separate complementation groups. These mutants exhibited the accumulation of poly A$\^$+/ RNA in the nucleus, with a decrease in the cytoplasm under synthetically lethal conditions, suggesting that the mutations cause an mRNA nuclear export defect. In addition, the S. pombe genes that were found to be involved in mRNA export did not suppress the synthetic lethality of these mutants. These results indicate that the isolated mutants contain mutations in new genes, which are involved in mRNA export from the nucleus.

Schizosaccharomyces pombe rsml Genetically Interacts with spmex67, Which Is Involved in mRNA Export

  • Yoon, Jin-Ho
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.32-36
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    • 2004
  • We have previously isolated three synthetic lethal mutants from Schizosaccharomyces pombe in order to identify mutations in the genes that are functionally linked to spmex67 with respect to mRNA export. A novel rsm1 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex1. The rsml gene contains no introns and encodes a 296 amino-add-long protein with the RING finger domain, a C3HC4 in the N-terminal half. The Δrsm1 null mutant is viable, but it showed a slight poly(A)$\^$+/ RNA accumulation in the nucleus. Also, the combination of Δrsm1 and Δspmex67 mutations confers synthetic lethality that is accompanied by the severe poly(A)$\^$+/ RNA export defect. These results suggest that rsm1 is involved in mRNA export from the nucleus.

Novel approaches for generating and manipulating diploid strains of Chlamydomonas reinhardtii

  • Kariyawasam, Thamali;Joo, Sunjoo;Goodenough, Ursula;Lee, Jae-Hyeok
    • ALGAE
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    • v.34 no.1
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    • pp.35-43
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    • 2019
  • Genetic study of haploid organisms offers the advantage that mutant phenotypes are directly displayed, but has the disadvantage that strains carrying lethal mutations are not readily maintained. We describe an approach for generating and performing genetic analysis of diploid strains of Chlamydomonas reinhardtii, which is normally haploid. First protocol utilizes self-mating diploid strains that will facilitate the genetic analysis of recessive lethal mutations by offering a convenient way to produce homozygous diploids in a single mating. Second protocol is designed to reduce the chance of contamination and the accumulation of spontaneous mutations for long-term storage of mutant strains. Third protocol for inducing the meiotic program is also included to produce haploid mutant strains following tetraploid genetic analysis. We discuss implication of self-fertile strains for the future of Chlamydomonas research.

A Forward Genetic Approach for Analyzing the Mechanism of Resistance to the Anti-Cancer Drug, 5-Fluorouracil, Using Caenorhabditis elegans

  • Kim, Seongseop;Shim, Jaegal
    • Molecules and Cells
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    • v.25 no.1
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    • pp.119-123
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    • 2008
  • Pyrimidine antagonists including 5-Fluorouracil (5-FU) have been used in chemotherapy for cancer patients for over 40 years. 5-FU, especially, is a mainstay treatment for colorectal cancer. It is a pro-drug that is converted to the active drug via the nucleic acid biosynthetic pathway. The metabolites of 5-FU inhibit normal RNA and DNA function, and induce apoptosis of cancer cells. One of the major obstacles to successful chemotherapy is the resistance of cancer cells to anti-cancer drugs. Therefore, it is important to elucidate resistance mechanisms to improve the efficacy of chemotherapy. We have used C. elegans as a model system to investigate the mechanism of resistance to 5-FU, which induces germ cell death and inhibits larval development in C. elegans. We screened 5-FU resistant mutants no longer arrested as larvae by 5-FU. We obtained 18 mutants out of 72,000 F1 individuals screened, and mapped them into three complementation groups. We propose that C. elegans could be a useful model system for studying mechanisms of resistance to anti-cancer drugs.