Cloning of cDNA encoding a soybean allergen, Gly m Bd 28K.

A cDNA clone encoding a soybean allergen, Gly m Bd 28K, has been isolated. The clone has a 1567-bp cDNA insert with a 1419-bp open reading frame and a 148-bp 3'-untranslated region, followed by a polyadenylation tail. The open reading frame was shown to encode a polypeptide composed of 473 amino acids. The chemically determined amino acid sequences of the peptides obtained from the allergen, including its N-terminal peptide, were shown to be contained in the N-terminal region of the amino acid sequence deduced from the cDNA, showing that the first half of the cDNA encodes the allergen with a preceding segment of 21 amino acids. The peptide fragment including the allergen was expressed as a fusion protein with glutathione S-transferase in Escherichia coli and immunoblotted with the sera of soybean-sensitive patients and the monoclonal antibody against the allergen. Furthermore, homology analyses demonstrate that the polypeptide for the cDNA exhibits high homology with the MP27/MP32 proteins in pumpkin seeds and the carrot globulin-like protein. This finding suggests that the polypeptide may consist of a 21-amino acid segment as a part of the signal peptide and the proprotein, which may be converted to two mature proteins, Gly m Bd 28K and a 23-kDa protein, during the development of soybean cotyledons.

A case study on Salmonella enteritidis (SE) origin at three egg-laying farms and its control with an S. enteritidis bacterin.

In the early 1990s, three egg-laying farms (farms S, T, and B) were thought to have the possibility of Salmonella enteritidis (SE) contamination because positive liquid egg samples originated from those farms. The present study was therefore conducted. The first clarification for SE contamination was the study on the origin of SE contamination including its vertical transmission. The results of SE contamination profiling with dust and manure, food materials, dead embryos, and residual yolks on hatch day in regular monitoring over a few years were clearly negative. Therefore, we concluded the SE transmission/infection was attributed to horizontal infection in the egg-laying farms but not vertical transmission from parental stock, hatcheries, growth, or food materials during a 7-yr experimental period. Second, we attempted to clarify if administration of an SE bacterin (Layermune SE) to growth flocks for the egg-laying farms could reduce SE incidence in liquid egg samples from each egg-laying farm. In the first experiment, we compared SE incidence in liquid egg samples from vaccinated and nonvaccinated flocks (similar age flocks). SE incidence from vaccinated and nonvaccinated flocks showed negative and <2 most probable number (MPN)/100 ml for farm B, <2 and >1600 MPN/100 ml for farm S, and negative and >1600 MPN/100 ml for farm T, respectively. In the second experiment, we compared the SE isolation incidence in the liquid egg samples from nonvaccinated and newly replaced vaccinated flocks in the same chicken houses from each of the three egg-laying farms. SE incidence in the liquid egg samples was similar to that in the first experiment. Therefore, the SE bacterin may play an important role in reducing the SE incidence of liquid egg samples.