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51.
Expression of cDNA for batroxobin, a thrombin-like snake venom enzyme 总被引:11,自引:0,他引:11
The cloned cDNA for batroxobin has been expressed in E. coli. Batroxobin could only be obtained as intracellular aggregates of fusion proteins, fused with a small peptide. To obtain the mature batroxobin, the recognition sequence for thrombin was inserted between the peptide and the mature batroxobin. This fusion protein accumulated in an insoluble form and could easily be purified. After site-specific cleavage of the fusion protein with thrombin, recombinant batroxobin was isolated by preparative electrophoresis. Batroxobin with enzymatic activity was obtained by the refolding of recombinant batroxobin. 相似文献
52.
The egg of the ascidian Ciona savignyi is pinkish red with brownish myoplasm that contains the putative determinants responsible for differentiation of muscle cells. When dechorionated unfertilized eggs were centrifuged at moderate speed, eggs were divided into centripetal, small gray fragments and centrifugal, large red fragments. The former contained the female pronucleus and clear cytoplasm, while most of the latter was filled with yolk granules. An antibody raised against the myoplasm of C. intestinalis eggs extensively stained the cortical region of gray fragments, while the antibody stained only small regions of the red fragments. After insemination, both fragments cleaved and gave rise to partial embryos. When development of muscle and epidermal cells in the partial embryos was examined with specific antibodies, muscle development was conspicuous in gray partial embryos, while epidermal differentiation was extensive in red partial embryos. Furthermore, when expression of markers of differentiation was examined in cleavage-arrested gray and red fragments, the number of arrested gray fragments exhibiting the muscle marker was about three-fold greater than in controls. These results suggest that putative muscle determinants are concentrated into gray fragments. 相似文献
53.
Koh Iba Ken-Ichiro Takamiya Yoshihiro Toh Hikaru Satoh Mitsuo Nishimura 《Genesis (New York, N.Y. : 2000)》1991,12(5):342-348
The ability to form functionally active chloroplasts is determined at a certain early stage of leaf development in three non-allelic temperature-sensitive virescent mutants of rice. Temperature-shift analysis, together with anatomical observations, indicates that the intrinsic developmental signals of the virescent genes are expressed at the stage immediately following the formation of basic leaf structure, but just before the onset of leaf elongation. These signals control the expression of chloroplast-encoded genes but do not affect the subsequent morphological development of the leaf or the photo-regulation of the expression of nuclear genes encoding chloroplast proteins. 相似文献
54.
Computer-aided molecular modelling of the endothelin (ET-A) receptor antagonists, BQ-123 and BE-18257B, shows that they have very similar 3D structures. Parts of their 3D structures are also shown to match closely with that reported for residues 6-8 in endothelin-1. On the basis of these similarities (and with supporting evidence from literature data on endothelin structure-activity relationships) a structural determinant is proposed for ET-A receptor binding, and novel designs of peptide are suggested for providing more potent and selective ET-A receptor antagonists. 相似文献
55.
cDNA cloning of an extracellular dermal glycoprotein of carrot and its expression in response to wounding 总被引:2,自引:0,他引:2
Suspension-cultured cells of carrot (Daucus carota L.) synthesize and secrete a glycoprotein that is normally found only in dermal tissues (epidermis, endodermis and periderm). This protein, previously called GP57, is now referred to as EDGP (E xtracellular D ermal G lyco P rotein). We purified sufficient quantities of EDGP to obtain amino-acid sequences on two internal tryptic peptides and screened a cDNA library of young carrot roots with antiserum to EDGP and with oligonucleotides corresponding to the peptides. Here we report the derived amino-acid sequence of EDGP. Sequence comparisons show that it has 40% amino-acid sequence identity with 7S basic globulin, a protein that is released when soybean seeds are soaked in hot water for a few hours. We suggest that these two proteins belong to a new family of dermal proteins. As far as we know, this is the first reported derived amino-acid sequence for protein that is specific to the epidermis and other dermal tissues. The level of EDGP mRNA is low in dry seeds, but increases rapidly in growing seedlings as they develop dermal tissues. The level of mRNA is low in storage roots, but increases rapidly in response to wounding. The presence of EDGP in dermal tissues and its up-regulation in response to wounding indicate a role in the response of plants to biotic and-or abiotic stresses. An unusual feature of the amino-acid sequence of EDGP is that it contains a short motif, which is present at the active site of aspartyl proteases such as pepsin and chymosin.Abbreviations cDNA
copy DNA
- 2,4-D
2,4-dichlorophen-oxyacetic acid
- EDGP
extracellular dermal glycoprotein
- 7SBG
7S basic globulin
Supported by a contract from the United States Department of Energy (Energy Biosciences) (to M.J.C.) and a Grant-in-Aid for Special Research on Priority Areas (01660002, Cellular and Molecular Basis for Reproductive Processes in Plants) from the Ministry of Education, Science and Culture, and by the Fund from Basic Research Core System of Science and Technology Agency, Japan (to S.S.). 相似文献
56.
57.
Proteins and Carbohydrates in Xylem Sap from Squash Root 总被引:8,自引:0,他引:8
Satoh Shinobu; Iizuka Chika; Kikuchi Akira; Nakamura Norio; Fujii Tadashi 《Plant & cell physiology》1992,33(7):841-847
The xylem sap from squash roots was collected from the cut surfaceof stems, and the proteins and carbohydrates in the sap wereanalyzed. The sap contained 18.6 µg ml1 proteinand the major polypeptides were as follows: 1) two polypeptides,of 75 and 40 kDa, with high-mannose glycans, the levels of whichincreased for about 24 h after cutting and then decreased; 2)a 32-kDa polypeptide, which appeared soon after cutting, disappearedand then reappeared again 4864 h after cutting; and 3)a 19-kDa and a 14-kDa polypeptide, which were present constitutively.The carbohydrates contained in the xylem sap were fractionatedinto 80% ethanol-soluble and -insoluble material, and whichwere analyzed by high-performance liquid chromatography, gaschromatography and enzymatic mathods. The former fraction containedconsiderable amounts of myo-inositol and fructose as free sugarsand oligosaccharides composed mainly of galactose, arabinoseand glucose. The latter contained polysaccharides composed mainlyof uronic acids, galactose and arabinose. The possible significanceof these substances, which may mediate the interactions betweenthe root and the aerial organs, is discussed. (Received April 20, 1992; Accepted July 4, 1992) 相似文献
58.
The Level of Stromal ATP Regulates Translation of the D1 Protein in Isolated Chloroplasts 总被引:1,自引:0,他引:1
The synthesis of the D1 subunit of the reaction center of photosystemII is light-dependent in isolated chloroplasts. The mechanismof the regulation by light was analyzed using spinach chloroplasts.The light-regulated synthesis of the D1 protein was preventedby the addition of atrazine and the dependence on the concentrationof atrazine of the inhibition was practically identical withthat of the inhibition of photosynthetic electron transportin photosystem II, as measured by the photoreduction of 2,6-dichlorophenolindophenol. Inhibitors of photosynthetic phosphorylation, suchas phloridzin, nigericin and carbonyl cyanide m-chlorophenylhydrazone,also inhibited the light-dependent synthesis of the D1 protein.Determination of the levels of ATP in chloroplasts and the ratesof synthesis of D1 protein under the various degrees of inhibitioncaused by these reagents suggested that the level of ATP inthe soluble, stromal fraction can control the synthesis of theD1 protein. The level of stromal ATP in chloroplasts was furthermanipulated, either by modulating the intensity of actinic lightor by the addition of metabolites, such as glycerate, whichwas used to decrease the level of ATP in the light, and dihydroxyacetonephosphate/oxaloacetate, which was used to raise the level ofATP in the dark. The results definitely support the hypothesisthat the light-induced level of ATP is an essential determinantin the regulation of the synthesis of the D1 protein in isolatedchloroplasts. (Received July 25, 1991; Accepted October 22, 1991) 相似文献
59.
60.
Effects of kainic acid on messenger RNA levels of IL-1 beta, IL-6, TNF alpha and LIF in the rat brain. 总被引:11,自引:0,他引:11
M Minami Y Kuraishi M Satoh 《Biochemical and biophysical research communications》1991,176(2):593-598
We examined the kainic acid-induced changes of mRNA levels of several cytokines such as IL-1 beta, IL-6, TNF alpha and LIF in the rat brain regions using semiquantitative RT-PCR method. IL-1 beta mRNA was markedly increased in the cerebral cortex (CC), thalamus (THL) and hypothalamus (HT) 2 h after the injection of kainic acid in a convulsive dose (12 mg/kg i.p.), and tended to decrease 4 h after the injection. IL-6 mRNA was weakly induced in the hippocampus (HPP) 2 h after the injection of kainic acid and was markedly increased in the CC, HPP, THL, and HT at 4 h. The level of TNF alpha mRNA was highly elevated in the CC, HPP, striatum (STR), THL and HT at 2 and 4 h after the injection. LIF mRNA apparently expressed in the CC and HPP of control rats and was increased in the CC, HPP and HT by the treatment with kainic acid. These results indicate that mRNAs of several cytokines are increased in various brain regions with different time-courses by kainic acid. 相似文献