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31.
Characterization of RNA binding specificity of the Drosophila sex-lethal protein by in vitro ligand selection. 总被引:8,自引:2,他引:6 下载免费PDF全文
The Drosophila sex-lethal (Sxl) protein, a regulator of somatic sexual differentiation, is an RNA binding protein with two potential RNA recognition motifs (RRMs). It is thought to exert its function on splicing by binding to specific RNA sequences within Sxl and transformer (tra) pre-mRNAs. To examine the Sxl RNA binding specificity in detail, we performed in vitro selection and amplification of ligand RNAs from a random sequence pool on the basis of affinity with Sxl protein. After three cycles of selection and amplification, we cloned and sequenced 17 cDNAs corresponding to the RNAs selected in vitro. Sequencing showed that most of the RNAs selected contain polyuridine stretches surrounded by purine residues. In vitro binding analysis revealed that the sequences of the in vitro selected RNAs with relatively high affinity for Sxl show similarity to that of the Sxl- and tra-regulated acceptor regions, including the invariant AG sequence for splicing. These results suggest that Sxl recognizes and preferentially binds to a polyuridine stretch with a downstream AG sequence. 相似文献
32.
Sakamoto Atsushi; Ohsuga Hiroyuki; Wakaura Makoto; Mitsukawa Norihiro; Hibino Takashi; Masumura Takehiro; Sasaki Yukiko; Tanaka Kunisuke 《Plant & cell physiology》1993,34(6):965-968
A cDNA clone for copper/zinc-superoxide dismutase (Cu/Zn-SOD)was isolated from spinach (Spinacia oleracea L.) leaves. Itsnucleotide sequence showed that it codes for a precursor polypeptideof 222 amino acids, including the NH2-terminal 68-residue extensionwhich corresponds to a plastidic transit peptide. Northern hybridization,using plastidic and cytosolic Cu/Zn-SOD cDNAs as the probes,revealed that these two genes are differentially expressed inthe roots and leaves of spinach.
1Present address: Department of Biochemistry and Microbiology,Cook College, Rutgers University New Brunswick, NJ 08903-0231,U.S.A. 相似文献
33.
Norio Tada Takuya Sakamoto Akihiko Kagami Keiko Mochizuki Kosei Kurosaka 《Molecular and cellular biochemistry》1993,119(1-2):171-178
Human plasmain vitro inhibits the growth of coagulase negative staphylococci,S. epidermidis, which may be pathogenic in the immunocompromised host. To determine the antimicrobial components, serum was fractionated by column chromatography, which revealed that elution areas where lipoproteins can be yielded had high antimicrobial activity againstS. epidermidis. Therefore, lipoprotein fractions, including very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL), were separated by ultracentrifugation and incubated withS. epidermidis. All 3 lipoprotein fractions suppressed bacterial growth within the first 3 h but VLDL enhanced bacterial growth after 9 h of incubation compared with the control. HDL, however, inhibited bacterial growth throughout 21 h of incubation.To confirm these results, serum from healthy volunteers was separated by ion exchange column chromatography and again by HPLC to purify the antimicrobial fraction. In the protein analysis with gradient polyacrylamide-SDS gel, apolipoprotein Al (apo Al), which is a major apolipoprotein of HDL, was detected in the antimicrobial fraction. Therefore, this fraction was loaded onto an immunoaffinity column coupled with the anti-apo Al monoclonal antibody (Mab). Unbound fraction had no antimicrobial activity, but anti-S. epidermidis activity was recovered from the bound fraction which consisted mainly of apo Al, All and apo C in protein composition.These results indicated that the antimicrobial activity was associated with the apo Al-containing lipoprotein particles (HDL). This property of HDL may directly affect bacterial growth and promote the self-defense mechanisms of normal and immunocompromised individuals. 相似文献
34.
Kazuki Sugawara Hidekazu Suzuki Mitsunobu Kamiya Keigo Osada Andrzej Witkowski 《Phycological Research》2023,71(4):182-192
The canal-bearing diatom genus Nagumoea, described based on only morphological evidence, was tentatively assigned to the order Bacillariales, although its phylogenetic position remained unclear. Because three isolates of Nagumoea (SK002, SK024 and SK053) were successfully established from Japanese coasts, we performed their morphological observations and molecular phylogenetic analyses to discuss the phylogeny and taxonomic position of this genus. Strains SK002 and SK024 were identified as Nagumoea africana, whereas SK053 conformed with Nagumoea serrata. There was high interspecific divergence between N. africana and N. serrata in the rbcL sequences (8.03–8.17%), indicating their distinctness. Furthermore, intraspecific variations were detected within N. africana (2.35%) in the rbcL, implying its cryptic diversity. The maximum likelihood and Bayesian phylogenetic trees inferred from the plastid rbcL, psbC and nuclear 18S rDNA genes recovered Nagumoea as monophyletic with strong statistical support and embedded within an unresolved, poorly supported lineage containing Achnanthes, Craspedostauros, Staurotropis and Undatella in the canal-bearing order Bacillariales (= the family Bacillariaceae). Although the constrained tree based on the monophyly of Nagumoea and the other canal-bearing clade (Surirellales and Rhopalodiales) was statistically rejected by the topology tests, the phylogenetic position of Nagumoea with other Bacillarialean members remains equivocal. The possession of two plastids positioned fore and aft, observed in the present study, and lack of keel, typical of the Bacillariales, indicate the possibility of Nagumoea being part of the ingroup of the Bacillariales or its closely related outgroup. 相似文献
35.
36.
Sakamoto Norihiro; Takagi Toshihisa; Sakaki Yoshiyuki 《Bioinformatics (Oxford, England)》1993,9(4):427-434
We have developed ODS (Overlapping Oligonucleotide Databasefor Signal Sequence Search)the first relational databasethat integrates information on biological features into thesearch for signal sequences. In existing biological sequencedatabases, even relational ones, retrieving nucleotide sequencesbased on their biological features involves much labour andtime or even the development of a new program. GenBank sequencedata, including FEATURES records, are organized into three relationaltables in ODS. Nucleotide sequences are transformed into overlappingoligonucleotides in order to facilitate the signal sequencesearch rapidly without the need for specific alignment programs.This transformation leads to a one-to-one correspondence betweenthe nucleotide sequence and its biological feature. The signalsequence search by ODS is done in SQL queries and ODS obviatesthe need for molecular biologists to write computer programs.The application of ODS to searches of promoter regions revealedputative cis-acting elements and basic statistical analysesof occurrences of oligonucleotides showed interesting findingsconcerning the cg dinucleotide. 相似文献
37.
Kazuo Sakamoto Kazuhiro Fujita Kimiko Hirayae Kumiko Iida Takashi Koyano Yoshihisa Asada Tsutomu Furuya 《Biotechnology Techniques》1993,7(10):749-754
Summary An integrated system has been constructed to instantly identify and efficiently sort the heterokaryons formed by plant protoplast fusion. The system is composed of the following functions: a) a transport system, b) an electro-manipulator, c) a cell harvester, d) a flow cytometer/cell sorter, and e) a control device. The conditions for an efficient and reproducible enrichment of the heterokaryons have been investigated by this system using the fluorescein isothiocyanate stained protoplasts preparing from Glycyrrhiza glabra cell cultures and unstained protoplasts of Abrus precatorius cell cultures which contain a large quantity of chlorophyll.Abbreviations MS
Murashige and Skoog (1962)
- 2,4-D
2, 4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- BAP
6-benzylaminopurine
- ABA
abscissic acid
- FITC
fluorescein isothiocyanate
This paper is part 96 in the series Studies on Plant Tissue Cultures. For part 95 see Orinara Y., Noguchi T. and Furuya T. (1993) submitted for publication. 相似文献
38.
Yaoyu Ren Timo Danner Alexandra Moy Martin Finsterbusch Tanner Hamann Jan Dippell Till Fuchs Marius Müller Ricky Hoft André Weber Larry A. Curtiss Peter Zapol Matthew Klenk Anh T. Ngo Pallab Barai Brandon C. Wood Rongpei Shi Liwen F. Wan Tae Wook Heo Martin Engels Jagjit Nanda Felix H. Richter Arnulf Latz Venkat Srinivasan Jürgen Janek Jeff Sakamoto Eric D. Wachsman Dina Fattakhova-Rohlfing 《Liver Transplantation》2023,13(1):2201939
The garnet-type phase Li7La3Zr2O12 (LLZO) attracts significant attention as an oxide solid electrolyte to enable safe and robust solid-state batteries (SSBs) with potentially high energy density. However, while significant progress has been made in demonstrating compatibility with Li metal, integrating LLZO into composite cathodes remains a challenge. The current perspective focuses on the critical issues that need to be addressed to achieve the ultimate goal of an all-solid-state LLZO-based battery that delivers safety, durability, and pack-level performance characteristics that are unobtainable with state-of-the-art Li-ion batteries. This perspective complements existing reviews of solid/solid interfaces with more emphasis on understanding numerous homo- and heteroionic interfaces in a pure oxide-based SSB and the various phenomena that accompany the evolution of the chemical, electrochemical, structural, morphological, and mechanical properties of those interfaces during processing and operation. Finally, the insights gained from a comprehensive literature survey of LLZO–cathode interfaces are used to guide efforts for the development of LLZO-based SSBs. 相似文献
39.
Nuclear localization activity of phytochrome B 总被引:31,自引:8,他引:23
Phytochromes are soluble red/far-red-light photoreceptor proteins which mediate various photomorphogenic responses of plants. Despite much effort, the signal transduction mechanism of phytochrome has remained obscure. Phytochromes are encoded by a small multigene family in Arabidopsis . Among the members of the family, phytochrome A (phyA) and B (phyB) are the best characterized. PhyB contains putative nuclear localization signals within its C-terminal region. Transgenic Arabidopsis plants were produced which expressed a fusion protein consisting of GUS and C-terminal fragments of phyB. GUS staining from the fusion protein in these transgenic plants was observed in the nucleus, which suggests that the nuclear localization signal of the fragment is functional. Next, it was examined whether the endogenous phyB was detected in the nucleus. Nuclei were isolated from the light-grown wild-type Arabidopsis leaves and subjected to the immunoblot analysis. The result indicated that a substantial fraction of total phyB was recovered in the isolated nuclei. This result was further confirmed by the immunocytochemical analysis of the protoplasts. Finally, the effects of light treatments on the levels of phyB in the isolated nuclei were examined. Dark adaptation of the plants before the nuclear isolation reduced the levels of phyB. The reduction was accelerated by irradiation of plants with far-red light before the transfer to darkness. Thus, nuclear localization of phyB was suggested to be light-dependent. 相似文献
40.
Miyake Ken; Ohtomi Michiko; Yoshizawa Hisamitsu; Sakamoto Yohko; Nakayama Katsumi; Okada Mitsumasa 《Plant & cell physiology》1995,36(1):109-113
Several water-soluble pigments were purified from gametangiaof Bryopsis maxima by liquid chromatography and characterizedby pyridylamination and high-performance anion-exchange chromatography.The structure of the main red pigment is proposed based on thedata of infrared spectrum, Mass spectrum, 1H and 13C NMR spectraand pyridylamino analysis. As a consequence, this pigment containeda tetrapyrrole with phytol and a sugar chain comprised of xyloseand glucose. The sequence of the sugars in the chain was determinedbased on its Mass spectrum. The pigment was similar to chlorophyll-originpigments observed in other plants. No aldehyde group, however,was present at C5 in the open tetrapyrrole chain. (Received August 3, 1994; Accepted November 10, 1994) 相似文献