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91.
Peggy Keeling Keith Johnson Daryl Sas Kathleen Klukas Peter Donahue Ross Johnson 《The Journal of membrane biology》1983,74(3):217-228
Summary The major membrane protein of the bovine lens fiber cell is a 26-kilodalton (kD) protein (MP26), which appears to be a component
of the extensive junctional specializations found in these cells. To examine the arrangement of MP26 within the junctional
membranes, various proteases were incubated with fiber cell membranes that had been isolated with or without urea and/or detergents.
These membranes were analyzed with electron microscopy and SDS-PAGE to determine whether the junctional specializations or
the proteins were altered by proteolysis. Microscopy revealed no obvious structural changes. Electrophoresis showed that chymotrypsin,
papain, and trypsin degraded MP26 to 21–22 kD species. A variety of protease treatments, including overnight digestions, failed
to generate additional proteolysis. Regions on MP26 which were sensitive to these three proteases overlapped. Smaller peptides
were cleaved from MP26 with V8 protease and carboxypptidases A and B. Protein domains cleaved by these proteases also overlapped
with regions sensitive to chymotrypsin, papain, and trypsin. Specific inhibition of the carboxypeptidases suggested that cleavage
obtained with these preparations was not likely due to contaminating endoproteases. Since antibodies are not thought to readily
penetrate the 2–3 nm extracellular gap in the fiber cell junctions, antibodies to MP26 were used to analyze the location of
the protease-sensitive domains. Antisera were applied to control (26 kD) and proteolyzed (22 kD) membranes, with binding being
evaluated by means of ELISA reactions on intact membranes. Antibody labeling was also done following SDS-PAGE and transfer
to derivatized paper. Both assays showed a significant decrease in binding following proteolysis, with the 22 kD product showing
no reaction with the anti-MP26 sera. These investigations suggest that MP26 is arranged with approximately fourfifths of the
primary sequence “protected” by the lipid bilayer and the narrow extracellular gap. One-fifth of the molecule, including the
C-terminus, appears to be exposed on the cytoplasmic side of the membrane. 相似文献
92.
Mathias Sprinzl Karl-Heinz Scheit Hans Sternbach Friedrich von der Haar Friedrich Cramer 《Biochemical and biophysical research communications》1973,51(4):881-887
2′-Deoxyadenosine and 3′-deoxyadenosine (cordycepin) can be incorporated into the 3′-terminal position of tRNAPhe by tRNA nucleotidyl transferase. tRNAPhe-C-C-2′dA and tRNAPhe-C-C-3′dA, missing the cis-diol group at the 3′-terminal end are resistant to periodate oxidation and are not able to form borate complexes. In aminoacylation experiments only the tRNAPhe-C-C-3′dA proved to be chargeable. 相似文献
93.
鉴定了小伞山羊草(Ae.umbellulata)6条染色体的中国春添加系对T型细胞质雄性不育系育性的影响,发现UAD能较好地恢复T型不育系的育性,表明染色体A上携带有育性恢复基因。添加染色体A在提莫菲维细胞质背景中通过雄配子的传递率为15.6%。同时进一步证明中国春不含有恢复基因。 在体细胞染色体数为42的331个不育系与UAD的杂种衍生后代中选到18个可育株,并对部分植株进行了细胞学鉴定。其中040-5、061-1和061-4与中国春的杂种F_1的育性分离和染色体配对情况表明它们是含有来自小伞山羊草染色体A上的恢复基因的杂合易位系。 相似文献
94.
J. J. M. Meulenberg W. A. M. Loenen E. Sellink P. W. Postma 《Molecular & general genetics : MGG》1990,220(3):481-484
Summary A generally applicable method is described for reintroduction of mutant plasmid-borne alleles to the chromosome of Klebsiella pneumoniae using bacteriophage . We, used this method to make stable chromosomal transposon insertions in genes for biosynthesis of pyrroloquinoline quinone in K. pneumoniae 相似文献
95.
Lisette Waits Stephanie Dunkle F. E. Wilkinson P. Moreau Keri Safranski T. Reust Dorothy M. Morré D. J. Morré 《Protoplasma》1990,154(1):8-15
Summary Transfer of radiolabeled lipids from dictyosome-like structures (DLS) from testis tubules of the guinea pig as donor to unlabeled plasma membrane from testis tubules immobilized on nitrocellulose as acceptor was studied in a completely cell-free system. As a general label for lipids of the donor DLS, isolated testis tubules were incubated with [14C]acetate. Time- and temperature-dependent transfer of [14C]acetate labeled constituents was observed in the cellfree system. However, despite the fact that phospholipids and other constituents were highly labeled in the donor fraction, primarily radioactive sterols were transferred to the plasma membrane acceptor vesicles. Transfer at 37°C represented 0.4 to 0.7% of the total radiolabeled cholesterol at 37°C but little or no transfer occurred at 4°C. The sterols transferred exhibited Chromatographic mobilities corresponding to those of cholesterol and lanosterol. Similar results were obtained with [14C]mevalonic acid. In subsequent experiments, cholesterol transfer from DLS to plasma membrane was demonstrated by incubation of DLS with [3H]squalene which was converted into sterol or with [14C]cholesterol. Transfer of sterols required ATP, but not cytosol, and was both time- and temperature-dependent. DLS were more effective than either endoplasmic reticulum or plasma membrane as the donor fraction. The results from the cell-free analysis suggest a possible functional role of the DLS in sterol biogenesis and transfer to the plasma membrane during spermatid development.Abbreviations DLS
dictyosome-like structure(s)
- PBS
phosphatebuffered saline
- HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- BSA
bovine serum albumin 相似文献
96.
F. Köhler I. Benediktsson G. Cardon C. S. Andreo O. Schieder 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,79(5):679-685
Summary In P. hybrida and B. nigra an enhancement of transformation rates (direct gene transfer) of about six to seven-fold was obtained after irradiation of protoplasts with 12.5 Gy (X-ray). The effect of protoplast irradiation was similar in experiments where protoplasts were irradiated 1h before transformation (X-ray/DNA) or 1h after completion of the transformation procedure (DNA/X-ray). Increased X-ray doses up to 62.5 Gy resulted in further enhancement of percentages of transformed colonies, indicating a correlation between relative transformation frequencies (RTF) and the doses applied. Estimation of degradation rates of plasmid sequences in plant protoplasts yielded a reduction of plasmid concentration to 50% 8–12 h after transformation. In 1-day-old protoplasts, the level of plasmid fragments dropped to 0%–10% compared to 1h after transformation. The results demonstrate that the integration rates of plasmid sequences into the plant genome may in part be governed by DNA repair mechanisms. This could be an explanation for the observed genotypic dependence of transformation rates in different plant species and plant genotypes. Gene copy number reconstructions revealed enhanced integration rates of plasmid sequences in transformed colonies derived from irradiated protoplasts. 相似文献
97.
T. Sakai J. Imamura 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,80(3):421-427
Summary Cytoplasts isolated from hypocotyl protoplasts of Raphanus sativus cv Kosena (cms line) by ultracentrifugation through Percoll/mannitol discontinuous gradient were fused with iodoacetamide(IOA)-treated protoplasts of Brassica napus cv Westar. Seventeen randomly selected regenerated plants were characterized for morphology and chromosome numbers. All of the regenerated plants had morphology identical to B. napus and 10 of them possessed the diploid chromosome number of B. napus. The remaining plants had chimeric or aneuploid chromosome numbers. The mitochondrial genomes in the 10 fusion products possessing the diploid chromosome numbers of B. napus were examined by Southern hybridization analysis. Four of the 10 plants contained mitochondrial DNA showing novel hybridization patterns. Of these 4 plants, 1 was male sterile, and 3 were male fertile. The remaining plants showed mitochondrial DNA patterns identical to B. napus and were male fertile. 相似文献
98.
Transfer of hygromycin resistance into Brassica napus using total DNA of a transgenic B. nigra line 总被引:3,自引:0,他引:3
The successful transfer of a marker gene (hpt gene) from Brassica nigra into B. napus via direct gene transfer was demonstrated. Total DNA was isolated from a hygromycin-resistant callus line, which contained three to five copies of the hpt gene. This line had been produced via direct gene transfer with the hygromycin resistance-conferring plasmid pGL2. The treatment of B. napus protoplasts with genomic DNA of B. nigra (HygR) resulted in relative transformation frequencies of 0.1–0.4%. Similar transformation rates were obtained in direct gene transfer experiments using B. napus protoplasts and plasmid pGL2. 相似文献
99.
László E. Heszky Zsolt Jekkel Abdel-Hamid Ali 《Plant Cell, Tissue and Organ Culture》1990,21(3):217-226
Reflexed saltmarsh-grass suspension cultures produced by seed callus were frozen to the liquid nitrogen temperature. Cooling rates, cryoprotectants and holding times were taken as a function of transfer temperatures. The highest survival of cells (45%) was found at a freezing rate of 1°C min-1, without cryoprotectant treatments. The cryoprotectants (proline, dimethyl sulphoxide, glycerol), used at different concentrations and transfer temperatures, increased the survival rate. The maximum value was 78% at 12.5% (w/v) of proline with –30°C transfer temperature. Considerable improvement of viability (from 0% to 95%) among the 12.5 and 15.0% (v/v) dimethyl sulphoxide cryopreserved cells was achieved by holding them at – 20°C for 10–30 min before plunging into the liquid nitrogen. A 20 min holding time at 15.0% (v/v) glycerol level and – 30°C transfer temperature significantly enhanced the viability of the explants from 42% to 92%. Plants were successfully regenerated from cells cryopreserved with proline (w/v) and dimethyl sulfoxide (v/v) levels of 12.5 and 15.0%, respectively. 相似文献
100.
The photosynthetic energy storage yield of uncoupled thylakoid membranes was monitored by photoacoustic spectroscopy at various measuring beam intensities. The energy storage rate as evaluated by the half-saturation measuring beam intensity (i50) was inhibited by 3-(3,4-dichlorophenyl)-1,1 dimethylurea, by heat inactivation or by artificial electron acceptors specific for photosystem I or photosystem II; and was activated by electron donors to photosystem I. The reactions involving both photosystems were all characterized by a similar maximal energy storage yield of 16±2 percent. The data could be interpreted if we assumed that the energy storage elicited by the photosystems at 35 Hz is detected at the level of the plastoquinone pool.Abbreviations PS
photosystem
- Tes
N-Tris [hydroxymethl] methyl-2-aminoethanesulfonic acid
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- DCIP
2,6-dichlorophenolindophenol
- FeCN
potassium ferricyanide
- DCBQ
2,5-dichlorobenzoquinone
- TMPD
N,N,N-tetramethyl-p-phenilenediamine 相似文献