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1.
Background
Protein translocation across the membrane of the Endoplasmic Reticulum (ER) is the first step in the biogenesis of secretory and membrane proteins. Proteins enter the ER by the Sec61 translocon, a proteinaceous channel composed of three subunits, α, β and γ. While it is known that Sec61α forms the actual channel, the function of the other two subunits remains to be characterized.Results
In the present study we have investigated the function of Sec61β in Drosophila melanogaster. We describe its role in the plasma membrane traffic of Gurken, the ligand for the Epidermal Growth Factor (EGF) receptor in the oocyte. Germline clones of the mutant allele of Sec61β show normal translocation of Gurken into the ER and transport to the Golgi complex, but further traffic to the plasma membrane is impeded. The defect in plasma membrane traffic due to absence of Sec61β is specific for Gurken and is not due to a general trafficking defect.Conclusion
Based on our study we conclude that Sec61β, which is part of the ER protein translocation channel affects a post-ER step during Gurken trafficking to the plasma membrane. We propose an additional role of Sec61β beyond protein translocation into the ER. 相似文献2.
Sequence of a cDNA for mouse thymidylate synthase reveals striking similarity with the prokaryotic enzyme 总被引:11,自引:0,他引:11
Perryman SM; Rossana C; Deng TL; Vanin EF; Johnson LF 《Molecular biology and evolution》1986,3(4):313-321
We report the nucleotide sequence of a cloned cDNA, pMTS-3, that contains a
1-kb insert corresponding to mouse thymidylate synthase (E.C. 2.1.1.45).
The open reading frame of 921 nucleotides from the first AUG to the
termination codon specifies a protein with a molecular mass of 34,962
daltons. The predicted amino acid sequence is 90% identical with that of
the human enzyme. The mouse sequence also has an extremely high degree of
similarity (as much as 55% identity) with prokaryotic thymidylate synthase
sequences, indicating that thymidylate synthase is among the most highly
conserved proteins studied to date. The similarity is especially pronounced
(as much as 80% identity) in the 44-amino-acid region encompassing the
binding site for deoxyuridylic acid. The cDNA sequence also suggests that
mouse thymidylate synthase mRNA lacks a 3' untranslated region, since the
termination codon, UAA, is followed immediately by a poly(A) segment.
相似文献
3.
Pelotti S Ceccardi S Alù M Lugaresi F Trane R Falconi M Bini C Cicognani A 《Genetic testing》2007,11(4):397-400
Microsatellites or short tandem repeats (STRs) markers are important tools for mapping disease-causing genes by linkage, for performing investigations in forensic medicine, for population genetic studies and for studying genetic modifications in tumors. In forensic applications neoplastic tissues can be used as a source of genetic information for personal identification or paternity testing when no other specimen is available. Cancer tissues can show microsatellite instability (MSI) and loss of heterozygosity (LOH) also for the STRs used in the forensic field. In this study, we screened 56 sporadic gastrointestinal carcinomas in order to provide further data for the evaluation of the incidence of allelic alterations for 15 STR loci and the suitability of using cancerous tissues in forensic applications. Sixty-six percent of the cancerous tissues were found to possess allelic alterations of the microsatellites analyzed with a high incidence of MSI-L (microsatellite instability low) when compared to the corresponding normal tissue. The most frequently altered loci were D18S51, VWA, and FGA. From a forensic perspective, great care must be taken in evaluating the DNA typing results obtained from cancerous tissue samples. 相似文献
4.
5.
The protein p12 accumulates in leaves of trees with citrus blight (CB), a serious decline of unknown cause. The function of p12 is not known, but sequence analysis indicates it may be related to expansins. In studies to determine the function of p12, sense and antisense constructs were used to make transgenic Carrizo citrange using an Agrobacterium-mediated transformation system. Homogeneous -glucuronidase+ (GUS+) sense and antisense transgenic shoots were regenerated using kanamycin as a selective agent. Twenty-five sense and 45 antisense transgenic shoots were in vivo grafted onto Carrizo citrange for further analyses. In addition, 20 sense and 18 antisense shoots were rooted. The homogeneous GUS+ plants contained either the p12 sense or antisense gene (without the intron associated with the gene in untransformed citrus) as shown by PCR and Southern blotting. Northern blots showed the expected RNA in the sense and antisense plants. A protein of identical size and immunoreactivity was observed in seven of nine sense plants but not in nine antisense or non-transgenic plants. At the current stage of growth, there are no visual phenotypic differences between the transgenic and non-transgenic plants. Selected plants will be budded with sweet orange for field evaluation for resistance or susceptibility to CB and general rootstock performance.Abbreviations AS
Acetosyringone
- CaMV 35S P
Cauliflower mosaic virus 35S promoter
- CaMV 35S poly A
Cauliflower mosaic virus 35S poly A terminator
- CB
Citrus blight
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- FMV
Figwort mosaic virus
- GUS
-Glucuronidase
- GUS gene
uidA
- IBA
Indole-3-butyric acid
- MES
2-(N-Morpholino) ethane sulfonic acid
- MSI
Inoculation medium
- MSP-10M
Plasmolysis solution with 10% maltose
- MSP-8S
Plasmolysis solution with 8% sucrose
- NAA
-Naphthaleneacetic acid
- NOS
Nopaline synthase
- NP
Nopaline synthase promoter
- NT
Nopaline synthase terminator
-
NPTII
Neomycin phosphotransferase II
- p12
Blight-associated protein p12 相似文献
6.
We have identified two single-copy genes from the model legume Medicago truncatula (MtENOD16 and 20) whose expression can be correlated with early stages of root nodulation and whose predicted coding sequences are partially homologous to both pea/vetch ENOD5 and soybean N315/ENOD55. Database searching and sequence alignment have defined the encoded early nodulins as a distinct sub-family of phytocyanin-related proteins, although the absence of key ligands implies that they are unlikely to bind copper. Molecular modelling based on known phytocyanin structure has been used to predict the 3-dimensional conformation of the principle globular domain of MtENOD16/20. Additional structural features common to both early nodulin and phytocyanin precursors include an N-terminal transit peptide, a highly variable (hydroxy)proline-rich sequence which probably undergoes extensive post-translational modification, and a hydrophobic C-terminal tail. 相似文献
7.
Brigida TL Lucena Billy M dos Santos João LS Moreira Ana Paula B Moreira Alvaro C Nunes Vasco Azevedo Anderson Miyoshi Fabiano L Thompson Marcos Antonio de MoraisJunior 《BMC microbiology》2010,10(1):298
Background
Bacteria may compete with yeast for nutrients during bioethanol production process, potentially causing economic losses. This is the first study aiming at the quantification and identification of Lactic Acid Bacteria (LAB) present in the bioethanol industrial processes in different distilleries of Brazil. 相似文献8.
9.
Role of the reticulum in the stability and shape of the isolated human erythrocyte membrane 总被引:5,自引:7,他引:5 下载免费PDF全文
In order to examine the widely held hypothesis that the reticulum of proteins which covers the cytoplamsic surface of the human erythrocyte membrane controls cell stability and shape, we have assessed some of its properties. The reticulum, freed of the bilayer by extraction with Triton X-100, was found to be mechanically stable at physiological ionic strength but physically unstable at low ionic strength. The reticulum broke down after a characteristic lag period which decreased 500-fold between 0 degrees and 37 degrees C. The release of polypeptide band 4.1 from the reticulum preceded that of spectrin and actin, suggesting that band 4.1 might stabilize the ensemble but is not essential to its integrity. The time-course of breakdown was similar for ghosts, the reticulum inside of ghosts, and the isolated reticulum. However, at very low ionic strength, the reticulum was less stable within the ghost than when free; at higher ionic strength, the reverse was true. Over a wide range of conditions the membrane broke down to vesicles just as the reticulum disintegrated, presumably because the bilayer was mechanically stabilized by this network. The volume of both ghosts and naked reticula varied inversely and reversibly with ionic strength. The volume of the naked reticulum varied far more widely than the ghost, suggesting that its deformation was normally limited by the less extensible bilayer. The contour of the isolated reticulum was discoid and often dimpled or indented, as visualized in the fluorescence microscope after labeling of the ghosts with fluoroscein isothiocyanate. Reticula derived from ghosts which had lost the ability to crenate in isotonic saline were shriveled, even though the bilayer was smooth and expanded. Conversly, ghosts crenated by dinitrophenol yielded smooth, expanded reticula. We conclude that the reticulum is a durable, flexible, and elastic network which assumes and stabilizes the contour of the membrane but is not responsible for its crenation. 相似文献
10.
Molecular evolution of a multigene family in group A streptococci 总被引:15,自引:0,他引:15
The emm genes are members of a gene family in group A streptococci (GAS)
that encode for antiphagocytic cell-surface proteins and/or
immunoglobulin-binding proteins. Previously sequenced genes in this family
have been named "emm," "fcrA," "enn," "arp," "protH," and "mrp"; herein
they will be referred to as the "emm gene family." The genes in the emm
family are located in a cluster occupying 3-6 kb between the genes mry and
scpA on the chromosome of Streptococcus pyogenes. Most GAS strains contain
one to three tandemly arranged copies of emm-family genes in the cluster,
but the alleles within the cluster vary among different strains.
Phylogenetic analysis of the conserved sequences at the 3' end of these
genes differentiates all known members of this family into four
evolutionarily distinct emm subfamilies. As a starting point to analyze how
the different subfamilies are related evolutionarily, the structure of the
emm chromosomal region was mapped in a number of diverse GAS strains by
using subfamily-specific primers in the polymerase chain reaction. Nine
distinct chromosomal patterns of the genes in the emm gene cluster were
found. These nine chromosomal patterns support a model for the evolution of
the emm gene family in which gene duplication followed by sequence
divergence resulted in the generation of four major-gene subfamilies in
this locus.
相似文献