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211.
Overproduction and nucleotide sequence of the respiratory D-lactate dehydrogenase of Escherichia coli. 总被引:5,自引:2,他引:3 下载免费PDF全文
Recombinant DNA plasmids containing the gene for the membrane-bound D-lactate dehydrogenase (D-LDH) of Escherichia coli linked to the promoter PL from lambda were constructed. After induction, the levels of D-LDH were elevated 300-fold over that of the wild type and amounted to 35% of the total cellular protein. The nucleotide sequence of the D-LDH gene was determined and shown to agree with the amino acid composition and the amino-terminal sequence of the purified enzyme. Removal of the amino-terminal formyl-Met from D-LDH was not inhibited in cells which contained these high levels of D-LDH. 相似文献
212.
A survey was done to find microorganisms useful for assaying sterigmatocystin; T-2 toxin and zearalenone.Staphylococcus aureus was found to be sensitive to T-2 toxin and zearalenone;Bacillus cereus was found to be sensitive to T-2 toxin only; andEscherichia coli was sensitive to sterigmatocystin. The response of the organisms to sterigmatocystin; T-2 toxin and zearalenone was found to be linear between 4 and 100 μg with sterigmatocystin toE. coli; between 2 and 25 μg with T-2 toxin toStaph, aureus andB. cereus; and between 4 and 100 μg with zearalenone toStaph, aureus. The lower limits of sensitivity of the test were 2 μg T-2 toxin and zearalenone, and 4 μg sterigmatocystin. The assay is rapid (15–17 hrs); simple and inexpensive; and can be used to verify the toxicity of samples and to confirm thin layer chromatographic results. 相似文献
213.
Evidence for effect of random genetic drift on G+C content after lateral transfer of fucose pathway genes to Escherichia coli K-12 总被引:4,自引:0,他引:4
The cps cluster of Escherichia coli K-12 comprises genes involved in
synthesis of capsular polysaccharide colanic acid. Part of the E. coli K-12
cps region has been cloned and sequenced and compared to its Salmonella
enterica LT2 counterpart. The cps genes from the two organisms are
homologous; in the case of the LT2 genes, with G+C content of 0.61 and
codons characteristic of high G+C species, it seems clear that they have
been acquired relatively recently by lateral transfer from a high G+C
species. The K-12 form of these cps genes is closely related to those of
LT2 so must derive from the same high G+C species, but it appears to have
transferred much earlier such that random genetic drift has brought P3 (the
corrected G+C content of codon base 3) down from 0.77 to 0.64, more than
halfway to the E. coli average of 0.57. We estimate, using an equation
developed by Sueoka, that the lateral transfer to E. coli took place
approximately 45 million years ago. This is the first report we are aware
of demonstrating the expected adjustment of P3 after lateral transfer
between species with different G+C content DNA.
相似文献
214.
Studies on the binding site of the galactose-specific agglutinin PA-IL from Pseudomonas aeruginosa 总被引:1,自引:0,他引:1
The binding properties of Pseudomonas aeruginosa agglutinin-I (PA-IL) with
glycoproteins (gps) and polysaccharides were studied by both the
biotin/avidin-mediated microtiter plate lectin-binding assay and the
inhibition of agglutinin-glycan interaction with sugar ligands. Among 36
glycans tested for binding, PA-IL reacted best with two glycoproteins
containing Galalpha1-->4Gal determinants and a human blood group ABO
precursor equivalent gp, but this lectin reacted weakly or not at all with
A and H active gps or sialylated gps. Among the mammalian disaccharides
tested by the inhibition assay, the human blood group Pkactive
Galalpha1-->4Gal, was the best. It was 7.4-fold less active than
melibiose (Galalpha1-->6Glc). PA-IL has a preference for the
alpha-anomer in decreasing order as follows: Galalpha1-->6
>Galalpha1-->4 >Galalpha1-->3. Of the monosaccharides studied,
the phenylbeta derivatives of Gal were much better inhibitors than the
methylbeta derivative, while only an insignificant difference was found
between the Galalpha anomer of methyl- and p -NO2-phenyl derivatives. From
these results, it can be concluded that the combining size of the
agglutinin is as large as a disaccharide of the alpha-anomer of Gal at
nonreducing end and most complementary to Galalpha1-->6Glc. As for the
combining site of PA-IL toward the beta-anomer, the size is assumed to be
less than that of Gal; carbon-6 in the pyranose form is essential, and
hydrophobic interaction is important for binding.
相似文献
215.
Moreno E; Lanne B; Vazquez AM; Kawashima I; Tai T; Fernandez LE; Karlsson KA; Angstrom J; Perez R 《Glycobiology》1998,8(7):695-705
P3 is a mouse monoclonal antibody (mAb) that binds to several NeuGc-
containing gangliosides. It also reacts with antigens expressed in human
breast tumors (Vazquez et al. (1995) Hybridoma , 14, 551-556). In this
work, the binding specificity of P3 has been characterized in more detail
using a panel of glycolipids that included several disialylated
gangliosides and several chemical derivatives of NeuGc-GM3. The carboxyl
group and the nitrogen function of sialic acid were found to play important
roles in the antibody binding, whereas the glycerol tail appears to be
nonrelevant. Molecular modeling was used to analyze the binding data,
including the finding that P3 selectively recognizes the internal NeuGc in
GD3. For this purpose, conformational studies of GD3 were performed using
molecular dynamics. It was concluded that sialic acid binds the P3 antibody
through its upper face (the one on which the carboxyl group is exposed) and
the C4-C5 side of the sugar ring, whereas none or very little contact
between the galactose residue and the protein is evident. Conformational
analysis of GD3 revealed that, despite the large flexibility of the
NeuGcalpha8NeuGc linkage, the P3 binding epitope on the external sialic
acid is not well exposed for any of the possible conformations this linkage
can adopt, whereas the internal sialic acid presents the epitope in a
proper way for several of these conformations. As a final result, a
coherent picture of the epitope that fits the wide binding data was
obtained.
相似文献
216.
The structural basis of molecular adaptation 总被引:31,自引:21,他引:10
The study of molecular adaptation has long been fraught with difficulties,
not the least of which is identifying out of hundreds of amino acid
replacements those few directly responsible for major adaptations. Six
studies are used to illustrate how phylogenies, site- directed mutagenesis,
and a knowledge of protein structure combine to provide much deeper
insights into the adaptive process than has hitherto been possible. Ancient
genes can be reconstructed, and the phenotypes can be compared to modern
proteins. Out of hundreds of amino acid replacements accumulated over
billions of years those few responsible for discriminating between
alternative substrates are identified. An amino acid replacement of modest
effect at the molecular level causes a dramatic expansion in an ecological
niche. These and other topics are creating the emerging field of
"paleomolecular biochemistry."
相似文献
217.
Marie-Elisabeth Samson-Bouma Nicole Verthier Leo A Ginsel Grard Feldmann Jack AM Fransen Lawrence P Aggerbeck 《Biology of the cell / under the auspices of the European Cell Biology Organization》1996,87(3):189-196
Summary— Intestinal biopsies from patients having genetic disorders of lipoprotein assembly and secretion, such as abetalipoproteinemia (ABL) or Anderson's disease (AD), contain large amounts of lipids which are accumulated in the enterocytes. Determination of the intracellular sites in which the lipids accumulate and to which apolipoproteins the lipids are bound would help to identify the defects in these diseases and further elucidate the mechanisms by which lipoprotein assembly and secretion occur normally. Ultrastructural immunogold labeling, however, is hampered by the poor preservation of the lipids accumulated in the enterocytes of these patients. We have used routine electron microscopy (fixation and ultra-thin sectioning) along with three methods for immunogold labeling of lipid-laden enterocytes; ultrathin cryosectioning, low temperature freeze substitution with embedding in Lowicryl K4M, and ultra-low temperature freeze substitution with embedding in Lowicryl HM20, to establish a protocol for investigating the intestinal tissue from these patients. Ultracryosectioning, while preserving the overall morphology of the lipid laden enterocytes, did not preserve the lipid content and the immunogold labeling of apolipoprotein B (ApoB) appeared dislocated. Freeze substitution and low temperature embedding in Lowicryl K4M, in contrast, appeared to better preserve the lipid and lipoprotein structures; however, the antigenicity of both apoAI and apoB appeared to be lost and no specific labeling could be obtained. Freeze substitution and embedding in Lowicryl HM20 best preserved the lipid and lipoprotein structures while maintaining apoprotein antigenicity. In conclusion, immunogold labeling of apolipoproteins on lipid structures in the lipid-laden enterocytes of patients with ABL and AD is best obtained by freeze substitution and embedding in Lowicryl HM20. 相似文献
218.
219.
Immunological similarities between specific chloroplast ribosomal proteins from Chlamydomonas reinhardtii and ribosomal proteins from Escherichia coli 总被引:11,自引:0,他引:11
Polyclonal antibodies were elicited against seven of the 33 different
proteins of the large subunit of the chloroplast ribosome from
Chlamydomonas reinhardtii. Three of these proteins are synthesized in the
chloroplast and four are made in the cytoplasm and imported. In western
blots, six of the seven antisera are monospecific for their respective
large subunit ribosomal proteins, and none of these antisera cross-reacted
with any chloroplast small subunit proteins from C. reinhardtii. Antisera
to the three chloroplast-synthesized ribosomal proteins cross-reacted with
specific Escherichia coli large subunit proteins of comparable charge and
molecular weight. Only one of the four antisera to the chloroplast
ribosomal proteins synthesized in the cytoplasm cross-reacted with an E.
coli large subunit protein. None of the antisera cross-reacted with any E.
coli small subunit proteins. On the assumption of a procaryotic,
endosymbiotic origin for the chloroplast, those chloroplast ribosomal
proteins still synthesized within the organelle appear to have retained
more antigenic sites in common with E. coli ribosomal proteins than have
those which are now the products of cytoplasmic protein synthesis. Antisera
to this cytoplasmically synthesized group of chloroplast ribosomal proteins
did not recognize any antigenic sites among C. reinhardtii cytoplasmic
ribosomal proteins, suggesting that the genes for the cytoplasmically
synthesized chloroplast ribosomal proteins either are not derived from the
cytoplasmic ribosomal protein genes or have evolved to a point where no
antigenic similarities remain.
相似文献
220.
Sterigmatocystin (STG) is a toxic metabolite produced by severalAspergillus species. Because of its toxic and carcinogenic properties the occurrence of STG in food is considered to represent a potential
hazard to man. The present study was designed to investigate following points:
相似文献
1 | A survey of STG incidence in Ras cheese on local markets. Ras cheese samples were collected from Cairo, Giza and Kalubia governorates. Thirty five percent of the samples contained the toxin with a mean value of 22.23 μg /kg |
2 | Fate of STG contaminating milk during Ras cheese processing. Milk was artificially contaminated with 125 μg/kg and processed into Ras cheese. Eighty percent of the toxin was distributed into the curd and 20% into the whey. Cheese ripening effected toxin content and the effect was temperature dependent. At 6°C: toxin concentration was slightly affected; at 20°C the toxin was reduced by 16% after 90 days when low toxin concentration was used. |
3 | Formation of STG byA versicolor mold on Ras cheese. Ras cheese blocks were contaminated with spores of the mold. Toxin production started after 45 days of ripening and reached a maximum at 90 days and then declined. Cow’s milk favoured toxin production over buffaloe’s. Aged cheese inhibited toxin production. |