ABSTRACT
SUMMARY: ASAP is a web tool designed to search for specific dipeptides, tripeptides and tetrapeptides in a protein sequence database. The server allows for: (a) identification of frequent and infrequent peptides and the creation of peptide probability tables for a given database of sequences (GenerNet program), (b) determination of the compatibility of an amino-acid sequence to the given peptide probability tables (ClonErrNet program); and (c) comparison of different protein databases based on peptide composition (CompNet program). ASAP server can be useful in protein engineering and/or protein classification studies.
Subject(s)
Oligopeptides/chemistry , Sequence Alignment/statistics & numerical data , Sequence Analysis, Protein/statistics & numerical data , Software , Computational Biology , Databases, Factual , Internet , Proteins/chemistryABSTRACT
Six fragments of the F gene from bovine respiratory syncytial virus (BRSV) were engineered into the pMAL-c2 Escherichia coli expression vector and expressed as C-terminal maltose-binding protein (MBP) fusion products. The resulting polypeptides were partially soluble and single-step purified by affinity chromatography. These fusion proteins were recognized in Western blots by several MAbs directed against human respiratory syncytial virus F protein. In addition, rabbit polyclonal antisera raised against two purified MBP-derived proteins reacted with the BRSV-F protein.
Subject(s)
Escherichia coli/genetics , HN Protein , Respiratory Syncytial Virus, Bovine/genetics , Viral Fusion Proteins/genetics , Viral Fusion Proteins/isolation & purification , Viral Proteins/biosynthesis , Viral Proteins/isolation & purification , Blotting, Western , Cloning, Molecular , DNA, Recombinant , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/isolation & purification , Respiratory Syncytial Virus, Bovine/chemistry , Solubility , Viral Envelope Proteins , Viral Fusion Proteins/biosynthesisABSTRACT
In order to identify the role of the bovine herpesvirus type 1 (BHV-1) glycoprotein E (gE) in the viral infection cycle, we have constructed a BHV-1 gE deletion mutant strain (BHV-1 gE-). This strain was assayed in vitro by comparing its growth kinetics with the wild type strain used as a host of the deletion. Our results indicate that those conditions which prevent the infection by direct adsorption to the cells (presence of a semi-solid medium or presence of neutralizing antibodies in the medium) selectively inhibit the growth of the gE- strain, suggesting that gE plays a central role in the BHV-1 spread by direct cell-to-cell transmission, a major mechanism of the BHV-1 in vivo virulence.
