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61.
Pairwise likelihood methods for inference in image models 总被引:3,自引:0,他引:3
62.
63.
MOTIVATION: Searching a protein sequence database for homologs is a
powerful tool for discovering the structure and function of a sequence. Two
new methods for searching sequence databases have recently been described:
Probabilistic Smith-Waterman (PSW), which is based on Hidden Markov models
for a single sequence using a standard scoring matrix, and a new version of
BLAST (WU-BLAST2), which uses Sum statistics for gapped alignments.
RESULTS: This paper compares and contrasts the effectiveness of these
methods with three older methods (Smith- Waterman: SSEARCH, FASTA and
BLASTP). The analysis indicates that the new methods are useful, and often
offer improved accuracy. These tools are compared using a curated (by Bill
Pearson) version of the annotated portion of PIR 39. Three different
statistical criteria are utilized: equivalence number, minimum errors and
the receiver operating characteristic. For complete-length protein query
sequences from large families, PSW's accuracy is superior to that of the
other methods, but its accuracy is poor when used with partial-length query
sequences. False negatives are twice as common as false positives
irrespective of the search methods if a family-specific threshold score
that minimizes the total number of errors (i.e. the most favorable
threshold score possible) is used. Thus, sensitivity, not selectivity, is
the major problem. Among the analyzed methods using default parameters, the
best accuracy was obtained from SSEARCH and PSW for complete-length
proteins, and the two BLAST programs, plus SSEARCH, for partial-length
proteins.
相似文献
64.
Paola Brusa Franco Dosio Francesca Pietribiasi Laura Delprino Paola Feraiorni Massimo Mariani Giovanni Bussolati Luigi Cattel 《Cancer immunology, immunotherapy : CII》1992,35(6):373-380
Summary We prepared a ricin-antibody conjugate, lacking the ability to bind the galactosidic residues of Sepharose 6B, a so-called blocked immunotoxin. The monoclonal antibody AR-3 was cross-linked to ricin through a thioether bond. Further studies showed that the immunoconjugate suppressed the tumour growth of HT-29 cells in intraperitoneally grafted nude mice, without showing any undesirable ricin toxicity.In this work, to demonstrate the therapeutic activity of the AR-3—ricin conjugate injected into mice bearing subcutaneous tumour, we first evaluated its pharmacokinetic behaviour and biodistribution. The behaviour of the immunoconjugate injected intravenously was almost intermediate between that of the antibody and ricin. Moreover, when the immunotoxin was intravenously administered to nude mice bearing subcutaneous tumour, no therapeutic effects appeared, in accordance with the relatively low permeability of the immunotoxin from the blood to the skin. In contrast, peritumoral treatment produced a strong reduction of the neoplastic nodules without substantial regrowth of the malignant cells. This result was also achieved when the immunotoxin treatment was performed on a well-established tumour. This finding was strictly related to the specificity of the immunoconjugate, since the analogous treatment with an irrelevant immunotoxin showed therapeutic failure. 相似文献
65.
66.
Recent horizontal transfer of a mariner transposable element among and between Diptera and Neuroptera 总被引:11,自引:4,他引:7
Transposable elements of the mariner family are widespread among insects
and other invertebrates, and initial analyses of their relationships
indicated frequent occurrence of horizontal transfers between hosts. A
specific PCR assay was used to screen for additional members of the
irritans subfamily of mariners in more than 400 arthropod species.
Phylogenetic analysis of cloned PCR fragments indicated that relatively
recent horizontal transfers had occurred into the lineages of a fruit fly
Drosophila ananassae, the horn fly Haematobia irritans, the African malaria
vector mosquito Anopheles gambiae, and a green lacewing Chrysoperla
plorabunda. Genomic dot-blot analysis revealed that the copy number in
these species varies widely, from about 17,000 copies in the horn fly to
three copies in D. ananassae. Multiple copies were sequenced from genomic
clones from each of these species and four others with related elements.
These sequences confirmed the PCR results, revealing extremely similar
elements in each of these four species (greater than 88% DNA and 95% amino
acid identity). In particular, the consensus sequence of the transposase
gene of the horn fly elements differs by just two base pairs out of 1,044
from that of the lacewing elements. The mosquito lineage has diverged from
the other Diptera for over 200 Myr, and the neuropteran last shared a
common ancestor with them more than 265 Myr ago, so this high similarity
implies that these transposons recently transferred horizontally into each
lineage. Their presence in only the closest relatives in at least the
lacewing lineage supports this hypothesis. Such horizontal transfers
provide an explanation for the evolutionary persistence and widespread
distribution of mariner transposons. We propose that the ability to
transfer horizontally to new hosts before extinction by mutation in the
current host constitutes the primary selective constraint maintaining the
sequence conservation of mariners and perhaps other DNA-mediated elements.
相似文献
67.
Luigi Cattel Laura Delprino Paola Brusa Franco Dosio Paolo M. Comoglio Maria Prat 《Cancer immunology, immunotherapy : CII》1988,27(3):233-240
Summary To avoid non-specific binding of intact ricin-antibody conjugates, we prepared a new blocked thioether-linkedricin-antibody IT, in which the galactosebinding site of ricin had lost the ability to bind to galactosidic residues of Sepharose 6B gel. As carrier agent, the monoclonal antibody AR-3, which defines the CAR-3 tumour-associated antigenic determinant expressed selectively on different human carcinoma cell lines, was used. Purification of the new conjugate was performed in three sequential steps: (1) by HPLC gel filtration on TSK G3000SW to remove the unconjugated ricin: (2) by affinity chromatography on Affi-Gel Blue to separate the free antibody from the conjugate and (3) by affinity chromatography on Sepharose 6B to separate the galactose-binding IT from the non-binding moiety. The cytotoxicity of the blocked and non-blocked thioether-linked IT was compared with that of classical ricin-antibody IT conjugated via SPDP and that of ricin A chain IT. The comparison was made on two different target cell lines (KATO III human gastric carcinoma and HT-29 human colorectal carcinoma) versus two control cell lines (HL-60 promyelocytic pre-leukaemic and COLO38 melanoma). The results showed that the blocked thioether IT displayed a more selective toxicity to target cells than the non-blocked IT and was much more potent than the ricin A chain conjugate.This work was supported by Grants from the Italian National Research Council (C.N.R. PFTBMS n. 86.01476.57 and n. 86.01748.03), the Ministry of Public Education and the Italian Association for Cancer Research (AIRC)
Abbreviations used: IT, immunotoxin; PBS, phosphate-buffered saline (100 mM sodium phosphate, 150 mM NaCl); SPDP, N-succinimidyl-3-(2-pyridyldithio)-propionate; PBS-EDTA, phosphatebuffered saline containing 1 mM EDTA; DMF, dimethylformamide; SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis; SIA, N-hydroxuccinimidyl ester of iodoacetic acid; SATA, N-succinimidyl-S-acetylthiolacetate 相似文献
68.
The effects of 18-crown-6 on the synthesis of peptides catalyzed by alpha-chymotrypsin are reported. Lyophilization of the enzyme in the presence of 50 equivalents of 18-crown-6 results in a 425-fold enhanced activity when the reaction between the 2-chloroethylester of N-acetyl-L-phenylalanine and L-phenylalaninamide is carried out in acetonitrile. Addition of crown ether renders the dipeptide synthesis in nonaqueous solvents catalyzed by alpha-chymotrypsin possible on a preparative scale. The acceleration is observed in different solvents and for various peptide precursors. Copyright 1998 John Wiley & Sons, Inc. 相似文献
69.
70.
Motomi Ito Kuniaki Watanabe Yoko Kita Takayuki Kawahara DJ Crawford Tetsukazu Yahara 《Journal of plant research》2000,113(1):79-89
Eupatorium were examined by sequencing the internal transcribed spacers (ITS) of nuclear ribosomal DNA and restriction site analysis
of chloroplast DNA. Molecular data provided strong evidence that (1) this genus originated in North America, (2) the genus
diverged into three morphological species groups, Eutrochium, Traganthes and Uncasia in North America, and (3) one of the North American Uncasia lineages migrated into temperate Europe and eastern Asia over the Bering land bridge. The estimated divergence times support
a late Miocene to early Pliocene migration from North America to Eurasia via the Bering land bridge. A European species was
sister to all of the eastern Asian species examined. The disjunct distribution pattern of the genus Eupatorium is incongruent with the classical Arcto-Tertiary geoflora concept.
Received 13 September 1999/ Accepted in revised form 4 January 2000 相似文献