@article {3437,
	title = {AQUEOUS 2-PHASE POLYMER SYSTEMS AS TOOLS FOR THE STUDY OF A RECOMBINANT SURFACE-EXPRESSED ESCHERICHIA-COLI HEMAGGLUTININ},
	journal = {Applied and Environmental Microbiology},
	volume = {61},
	number = {9},
	year = {1995},
	note = {ISI Document Delivery No.: RT798Times Cited: 5Cited Reference Count: 15},
	month = {Sep},
	pages = {3251-3255},
	type = {Article},
	abstract = {The surface expression of an integral membrane hemagglutinin, HRA1, cloned from Escherichia coli O9:H10:K99 in heterologous E. coli strains was studied by utilizing a variety of polyethylene glycol-dextran and dextran-Ficoll aqueous two-phase polymer systems. Bacteria containing plasmids that encoded the hemagglutinin were found to partition differently from both the host bacteria lacking the plasmid and the original hemagglutinating strain in several of these systems. By using molecular biological techniques, the origin of the partition difference was unambiguously correlated to the expression of HRA1, providing evidence independent of the agglutination phenotype that the protein was accessible to the surrounding milieu. It Was demonstrated by using bacterial partition in charge-sensitive systems that the agglutination event was not likely to be due to the presence of a nonspecific positively charged surface protein, as HRA1-expressing clones showed no less affinity for the relatively positive polyethylene glycol-rich upper phase than did control bacteria, This work demonstrates the utility of aqueous polymer two-phase systems for the study of surface-expressed recombinant proteins, due to the sensitivity of the systems and the presence of excellent controls (the host bacteria before plasmid introduction). In cloning and expression studies of surface-associated proteins, two-phase aqueous polymer systems could be used as an alternative to antibody production for the monitoring of surface expression, and these systems may give valuable information on the surface exposure of the protein.},
	keywords = {ANTIGEN, CLONING, PROTEIN, STRAINS},
	isbn = {0099-2240},
	url = {<Go to ISI>://A1995RT79800010},
	author = {Lutwyche, P. and Norrisjones, R. and Brooks, D. E.}
}
@article {3106,
	title = {CLONING, SEQUENCING, AND VISCOMETRIC ADHESION ANALYSIS OF HEAT-RESISTANT AGGLUTININ 1, AN INTEGRAL MEMBRANE HEMAGGLUTININ FROM ESCHERICHIA-COLI-O9-H10-K99},
	journal = {Infection and Immunity},
	volume = {62},
	number = {11},
	year = {1994},
	note = {ISI Document Delivery No.: PN304Times Cited: 13Cited Reference Count: 45},
	month = {Nov},
	pages = {5020-5026},
	type = {Article},
	abstract = {The gene encoding a mannose-resistant hemagglutinating protein was cloned from Escherichia coli 09:H10:K99. The hemagglutinin is different from two other mannose-resistant hemagglutinins in this strain, K99 and F41. The agglutinin, named heat-resistant agglutinin 1 (HRA1) since heating to 70 degrees C does not destroy its aggregative properties, strongly agglutinates human, pig, and dog erythrocytes, shows little or no affinity towards cow and chicken erythrocytes, but agglutinates human colon adenocarcinoma 201 (COLO 201) cells. The hra1 gene present on the recombinant plasmid pETE1 was localized by subcloning, and its nucleotide sequence was determined. The gene consists of a 792-bp open reading frame coding for a putative protein of 29 kDa with a predicted N-terminal secretory signal sequence. HRA1 shares no significant identity with data base protein sequences. HRA1 is strongly associated with the bacterial membrane, resisting sonication and isolation attempts based upon standard adhesin purification techniques. N-terminal sequencing of a unique 25-kDa band present in polyacrylamide gels of outer membrane preparations of bacteria harboring pETE1 correlated with the predicted N-terminal amino acid sequence of HRA1 after cleavage of the signal peptide. A viscometric agglutination assay sensitive to the strength of bacterial adhesion shows that the agglutination mediated by bacteria expressing HRA1 is weaker than that of bacteria bearing the F41 adhesin, probably because of the high-molecular-weight, multivalent nature of the latter adhesin. Our observations suggest that HRA1 is a monomeric outer membrane agglutinin.},
	keywords = {ANTIGEN, BACTERIA, DNA, K99, LOCALIZATION, NONFIMBRIAL, OUTER-MEMBRANE, PROTEINS, PURIFICATION, STRAINS},
	isbn = {0019-9567},
	url = {<Go to ISI>://A1994PN30400046},
	author = {Lutwyche, P. and Rupps, R. and Cavanagh, J. and Warren, R. A. J. and Brooks, D. E.}
}
