The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs.
AuthorsKwan KM, Fujimoto E, Grabher C, Mangum BD, Hardy ME, Campbell DS, Parant JM, Yost HJ, Kanki JP, Chien CB,
JournalDev Dyn
PubMed ID17937395
'Transgenesis is an important tool for assessing gene function. In zebrafish, transgenesis has suffered from three problems: the labor of building complex expression constructs using conventional subcloning; low transgenesis efficiency, leading to mosaicism in transient transgenics and infrequent germline incorporation; and difficulty in identifying germline integrations unless using a fluorescent ... More
Functional analysis of the ATG8 homologue Aoatg8 and role of autophagy in differentiation and germination in Aspergillus oryzae.
AuthorsKikuma T, Ohneda M, Arioka M, Kitamoto K,
JournalEukaryot Cell
PubMed ID16896216
'Autophagy is a well-known degradation system, induced by nutrient starvation, in which cytoplasmic components and organelles are digested via vacuoles/lysosomes. Recently, it was reported that autophagy is involved in the turnover of cellular components, development, differentiation, immune responses, protection against pathogens, and cell death. In this study, we isolated the ... More
Transcriptional and posttranscriptional regulation of transcription factor expression in Arabidopsis roots.
AuthorsLee JY, Colinas J, Wang JY, Mace D, Ohler U, Benfey PN,
JournalProc Natl Acad Sci U S A
PubMed ID16581911
'Understanding how the expression of transcription factor (TF) genes is modulated is essential for reconstructing gene regulatory networks. There is increasing evidence that sequences other than upstream noncoding can contribute to modulating gene expression, but how frequently they do so remains unclear. Here, we investigated the regulation of TFs expressed ... More
BopC is a novel type III effector secreted by Bordetella bronchiseptica and has a critical role in type III-dependent necrotic cell death.
AuthorsKuwae A, Matsuzawa T, Ishikawa N, Abe H, Nonaka T, Fukuda H, Imajoh-Ohmi S, Abe A,
JournalJ Biol Chem
PubMed ID16407269
'In Bordetella bronchiseptica, the functional type III secretion system (TTSS) is required for the induction of necrotic cell death in infected mammalian cells. To identify the factor(s) involved in necrotic cell death, type III-secreted proteins from B. bronchiseptica were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and electrospray ionization ... More
A novel high-throughput (HTP) cloning strategy for site-directed designed chimeragenesis and mutation using the Gateway cloning system.
AuthorsSuzuki Y, Kagawa N, Fujino T, Sumiya T, Andoh T, Ishikawa K, Kimura R, Kemmochi K, Ohta T, Tanaka S,
JournalNucleic Acids Res
PubMed ID16009811
There is an increasing demand for easy, high-throughput (HTP) methods for protein engineering to support advances in the development of structural biology, bioinformatics and drug design. Here, we describe an N- and C-terminal cloning method utilizing Gateway cloning technology that we have adopted for chimeric and mutant genes production as ... More
Caenorhabditis elegans reporter fusion genes generated by seamless modification of large genomic DNA clones.
AuthorsDolphin CT, Hope IA,
JournalNucleic Acids Res
PubMed ID16717278
By determining spatial-temporal expression patterns, reporter constructs provide significant insights into gene function. Although additionally providing information on subcellular distribution, translational reporters, where the reporter is fused to the gene coding sequence, are used less frequently than simpler constructs containing only putative promoter sequences. Because these latter constructs may not ... More
Development of a versatile expression plasmid construction system for Aspergillus oryzae and its application to visualization of mitochondria.
AuthorsMabashi Y, Kikuma T, Maruyama J, Arioka M, Kitamoto K,
JournalBiosci Biotechnol Biochem
PubMed ID16880596
We report here a development of the MultiSite Gateway(TM)-based versatile plasmid construction system applicable for the rapid and efficient preparation of Aspergillus oryzae expression plasmids. This system allows the simultaneous connection of the three DNA fragments inserted in entry clones along with a destination vector in a defined order and ... More
Membrane transporters in the relict plastid of malaria parasites.
AuthorsMullin KA, Lim L, Ralph SA, Spurck TP, Handman E, McFadden GI,
JournalProc Natl Acad Sci U S A
PubMed ID16760253
Malaria parasites contain a nonphotosynthetic plastid homologous to chloroplasts of plants. The parasite plastid synthesizes fatty acids, heme, iron sulfur clusters and isoprenoid precursors and is indispensable, making it an attractive target for antiparasite drugs. How parasite plastid biosynthetic pathways are fuelled in the absence of photosynthetic capture of energy ... More
Evidence for high specificity and efficiency of multiple recombination signals in mixed DNA cloning by the Multisite Gateway system.
AuthorsSasaki Y, Sone T, Yoshida S, Yahata K, Hotta J, Chesnut JD, Honda T, Imamoto F,
JournalJ Biotechnol
PubMed ID14736459
Six types of recombination signal DNA sequences of the Multisite Gateway cloning system were investigated as to their specificity and efficiency in the LR and BP recombination reactions. In the LR reaction to generate an Expression clone by recombination between attL and attR signals which are contained in the Entry ... More
Building blocks for plant gene assembly.
AuthorsKarimi M, Bleys A, Vanderhaeghen R, Hilson P,
JournalPlant Physiol
PubMed ID17965171
The MultiSite Gateway cloning system, based on site-specific recombination, enables the assembly of multiple DNA fragments in predefined order, orientation, and frame register. To streamline the construction of recombinant genes for functional analysis in plants, we have built a collection of 36 reference Gateway entry clones carrying promoters, terminators, and ... More