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201.
The genome of Saccharomyces cerevisiae contains five genes that encode type II transmembrane proteins with significant amino acid similarity to the alpha-1,3-mannosyltransferase Mnn1p. The roles of the three genes most closely related to MNN1 were examined in mutants carrying single and multiple combinations of the disrupted genes. Paper chromatographic analysis of [2-3H]mannose-labeled O-linked oligosaccharides released by beta-elimination showed that the MNT2 (YGL257c) and MNT3 (YIL014w) genes in combination with MNN1 have overlapping roles in the addition of the fourth and fifth alpha-1,3-linked mannose residues to form Man4 and Man5 oligosaccharides whereas MNT4 (YNR059w) does not appear to be required for O-glycan synthesis. 相似文献
202.
M McClarrinon L Gilkey V Watral B Fox C Bullock L Fradkin D Liggitt L Roche L B Bussey E Fox C Gorman 《DNA and cell biology》1999,18(7):533-547
As the sequencing of the human genome proceeds, the need for a new screen for in vivo function is becoming apparent. Many investigators are turning to various transgenic models as a means of studying function. However, these approaches are very time consuming, with a transgene-expressing mouse model often taking months to establish. We have developed an efficient system for delivering genes in vivo, which allows the gene product to be studied as early as 24 h after introduction into the mouse model. The delivery system employs a novel cationic lipid, 1-[2-(9-(Z)-octadecenoyloxy)ethyl]-2-(8-(Z)-heptadecenyl)-3- (hydroxyethyl)imidazolinium chloride (DOTIM), and a neutral lipid, cholesterol, complexed with an expression vector containing the reporter gene chloramphenicol acetyl transferase (CAT). After a single intravenous injection of these complexes, several tissues were seen to express the transgene. High, persistent expression in the vascular endothelial cells in the mouse lung was obtained. Delivery of DNA in vivo has been evaluated by quantitative polymerase chain reaction and protein expression by CAT activity assays. In vivo studies showed reproducible expression in more than 500 mice injected via the tail vein. An early peak of expression was followed by lower, but sustained, expression for > 50 days. Transgene expression of CAT could also be identified by immunohistochemistry staining in mouse lung and appeared to be located within the capillaries. The pattern of in vivo expression could be modulated and targeted to specific organs by altering the lipid-DNA formulation. New expression vectors with altered introns and polyadenylation sites further improved expression. The expression reported here may be sufficient in magnitude, duration, and flexibility to be an attractive alternative, in some cases, to establishing transgenic animals by stable gene transfer. 相似文献
203.
D T Rogers D Saville H Bussey 《Biochemical and biophysical research communications》1979,90(1):187-193
The extracellular proteins and glycoproteins of a yeast mutant defective in killer toxin expression were separated by one and two dimensional polyacyylamide gel electrophoresis. Many mutant extracellular proteins and glycoproteins show both altered electrophoretic mobility and isoelectric points when compared with the parent strain. Altered proteins and glycoproteins from were identified with their parental counterparts by peptide mapping. The observed alterations co-segregated with the nuclear mutation in genetic crosses. 相似文献
204.
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206.
Summary A uracil-sensitive mutant of Salmonella typhimurium was isolated by diethyl sulfate mutagenesis and penicillin counterselection. This mutation identifies a new Salmonella gene that is well separated from the structural genes for arginine and pyrimidine biosynthesis. The use-1 mutation was located between the ilv gene cluster (isoleucine-valine operon) and hisR (structural gene for histidine tRNA) at 83 map units. 相似文献
207.
Chromosomal mutants of Saccharomyces cerevisiae affecting the cell wall binding site for killer factor. 总被引:6,自引:0,他引:6
Fifty-two killer, factor-resistant, nuclear mutants were isolated from sensitive strains of yeast and assorted into three functional groups. All but one mutant owed their resistance to an alteration in the cell wall binding site for killer. In several mutant strains, an alteration at the site of killer binding was associated with a change in the susceptibility of the cell wall to degradation by glusulase. The killer-binding site could be inactivated by periodate but not by pronase treatment. The nature of the site is discussed. 相似文献
208.
K1 killer toxin, a pore-forming protein from yeast 总被引:21,自引:0,他引:21
H. Bussey 《Molecular microbiology》1991,5(10):2339-2343
K1 killer toxin is a secreted, pore-forming protein that kills sensitive yeast cells. The heterodimeric toxin is processed from a precursor in the Golgi, and has allowed identification of the KEX2- and KEX1-encoded proteases. The toxin binds to a glucan receptor on the cell wall of target yeast, and mutational analysis implicates both the alpha- and beta-toxin subunits in receptor binding. Toxin-resistant mutants with altered cell-wall glucans have helped to outline a pathway of assembly of these polysaccharides. Patch-clamp technology has demonstrated the nature of the lethal channel in toxin-treated plasma membranes. The hydrophobic alpha-subunit-encoding region is the site of all mutations affecting channel formation. Immunity to the toxin is conferred by the toxin precursor, and immunity mutations map to the region encoding the alpha subunit. The precursor probably competes with the toxin to prevent channel formation in toxin-producing cells, but the basis of this remains unknown. This toxin/immunity system has a domain structure that differs from that of other characterized toxins and has no known homologues. 相似文献