ISSN:0749-503X    

DOI:10.1002/yea.320060102

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractRecently a convenient method for detection of specific RNA sequences in bacteria has been developed (Ivanov and Gigova, 1986) but the original protocol was inapplicable to microorganisms with a rigid cell wall. Here we report a modification of the RNA colony hybridization for use with yeast. The modified method includes the following consecutive procedures: (a) treatment of the yeast colonies on the membrane filter with 10% SDS at 65°C for 30 min; (b) treatment of the same filter with 3 × SSC, 10% formaldehyde at 65°C for 30 min; (c) hybridization with32P‐labelled oligonucleotide or (DNA) specific for the RNA sequence of interest. The intensity of the radioactive signals thus obtained is comparable with that of theE. colsicolo

ISSN:0749-503X    

DOI:10.1002/yea.320060103

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractWe have the substructure and polypeptide composition of the peroxisomal membranes in two methylotrophic yeasts in relation to different growth conditions. The results obtained that no significant ultrastructural differences existed between the membranes of variously grown cells.The presence of specific peroxisomal membrane proteins (PMPs) was studied biochemically. On sodium dodecyl sulphate‐polyacrylamide gels of purified microbody membranes isolated from methanol‐grownHansenula polymorpha, prominent proteins bands were observed at 22, 31, 35, 42, 49 and 51 kD. These proteins were also present when the cells were grown in media containing ethanol and/or ethylamine. Apart from these, several other PMPs were specifically induced under these conditions, namely 24, 29, 37 and 62 kD proteins. The polypeptide composition of peroxisomal membranes fromH. polymorphawas compared with that of another methylotroph.Candida biodinii. In the latter organism a specific PMP with a molecular weight of 23 kD was induced during growth onD‐alanine instead of ammonium sulphate as the nitrogen s

ISSN:0749-503X    

DOI:10.1002/yea.320060104

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractThe presence of an ATpase on yeast peroxisomal membranes was studied by immunological methods. Western blot analysis of purified peroxisomal membranes from several yeasts revealed distinct cross‐reaction with specific antibodies against the F1‐part or the β‐subunit of the mitochondrial ATpase ofSaccharomyces cerevisiae. This was not due to mitochondrial contamination as was demonstrated by analytical sucrose gradient centrifugation.Protein A‐gold labelling carried out on Lowicryl‐embedded methanol‐grownHansenula polymorphausing these antibodies did not result in significant staining. However, when organelles isolated from this yeast were successively incubated with antibodies and protein A‐gold prior to embedding, specific labelling was observed on both the peroxisomal membrane and the membrane of damaged mitochondria but not on intact mitochondira. Specific labelling of the peroxisomal membrane was confirmed by freeze‐fracture immunocytochemistry. In addition to the peroxisomal membrane, the mitochondrial membrane was also labelled in these experiments. Freeze‐fracture immunocytochemistry was also successful for the localization of peroxisomal matrix proteins, e.g. alcohol oxidase and dihydroxyacetone synthase and of mitochondrial membrane proteins, e.g. cy

ISSN:0749-503X    

DOI:10.1002/yea.320060105

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractUsing the techniques of centrifugal elutriation it was demonstrated that during the cell division cycle of the budding yeastSaccharomyces cerevisiaethere are stage‐specific fluctuations in the intracellular concentration of adenosine 3′,5′‐cyclic monophosphate (cAMP). Results shown here indicate that the intracellular concentration of cAMP is at its highest during the division cycle, and its lowest immediately prior to and just after cell sepraration. Results also show the extrusion of extracellular cAMP into the medium bySaccharomyces cerevisiae, extracellular cAMP levels being ten to one hundred times higher than intracellular levels. During the cell ofSaccharomyces cerevisiaethe extracellular level of cAMP does not fl

ISSN:0749-503X    

DOI:10.1002/yea.320060106

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractThe occurrence of cytochrome P‐45014DM(lanosterol 14α‐demethylase) and cytochrome P‐450alk(long‐chain alkane terminal hydroxylase) in various yeast strain was determined with immunological procedures. Cytochrome P‐45014DM, which is a constitutive or housekeeping enzyme playing an essential role in ergosterol biogenesis, was found in all yeast strains so far tested. Cytochromes P‐45014DMfrom different species of yeast were immunologically different, although they may have a few common antigenic sites. In contrast, cytochrome P‐450alkwas detected only in the alkane‐ass

ISSN:0749-503X    

DOI:10.1002/yea.320060107

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractWe have undertaken a search for autonomously replicating (ARSs) fromKluvermyces lactischromosomal DNA able to sustain plasmid replication inK. lactisand inSaccharomyces cerevisiae. The discovery of such sequences might be interesting for the comparison of ARSs from different sources and possibly useful for the construction of multivalent vectors.HindIII fragments fromK. lactischromosomal DNA were inserted in the YIp5 plasmid (lacking an origin of replication) and the resulting chimaeric plasmids were selected for the ability to transformS. cerevisiae. Four plasmids were identified and further analysed. Two contained the same 1.8 kbK. lactisfragment and transformed bothK. lactisandS. cerevisiaewith the same efficiency and stability, whereas the third transformed onlyS. cerevisiaeand the fourth transformedK. lactiswith a higher efficiency thanS. cerevisiae.A detailed study was performed on the 1.8kb fragment which exhibited ARS function in both yeasts. The fragment was subcloned using different restriction enzymes andBal31 exonuclease. Subclones were tested for ARS function. ARS activities in the two yeasts were localized in the same 100 bp region. Sequencing demonstrated the presence in this region of the dodecanucleotide 5′ATTTATTGTTTT 3′ differing from the ARS core consensus ofS. cerevisiaeonly by a T insertion.A similar nucleotide sequence ofS. cerevisiaeonly in the putative replication origin of the 2 μ‐like plasmid pkD1 which stably replicates inK. lactis. Homologies with ARSs fromS. cerevisiaewere also found in the regions flanking the above‐mentioned dodecanu

ISSN:0749-503X    

DOI:10.1002/yea.320060108

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

摘要:

AbstractYeast flocculation was inhibited by high concentrations of a number of salts. Calcium and magnesiun salts were potent inhibitors and cesium salts were least effective. Partial inhibitors by different salts additive and were completely reversible by salt removal. Inhibition by salts was time dependent; prolonged incubation increased the degree of inhibition. Salt inhibition was partly caused by the action of salts lowering the buffer pH value, and partly caused by chaotropic inhibition of proteins on the surfaces of flocculent cells. Flocculation receptors on non‐flocculent cells were ubaffected by high salt concentration. At sub‐inhibitory salt concentrations, there was an enhancement of flocculation in both rate and ext

ISSN:0749-503X    

DOI:10.1002/yea.320060109

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY

ISSN:0749-503X    

DOI:10.1002/yea.320060101

出版商:John Wiley&Sons, Ltd.    

年代:1990

数据来源: WILEY