ABSTRACT
Protein I from the outer membrane of Escherichia coli and other members of the family Enterobacteriaceae is a potent mitogen and polyclonal B-lymphocyte activator. To determine the part of the polypeptide responsible for biological activity, we cleaved the molecule into defined polypeptide fragments of approximate molecular weights 24,000, 15,000, 9,000, 7,000, and 3,000 by using the cyanogen bromide method. The fragments were purified by gel permeation chromatography and by preparative polyacrylamide gel electrophoresis. They were investigated for mitogenicity and for the induction of immunoglobulin synthesis in lymphocyte cultures from several inbred mouse strains. The fragment of molecular weight 24,000 turned out to be a potent polyclonal B-lymphocyte activator comparable to native protein I. The low-molecular-weight fragments exhibited only marginal effects. Neither purified T lymphocytes nor thymocytes were activated. Our results show that a defined fragment of protein I is responsible for its lymphocyte-stimulating activity.
Subject(s)
Antigens, Bacterial/immunology , B-Lymphocytes/immunology , Bacterial Outer Membrane Proteins/immunology , Animals , Kinetics , Lymphocyte Activation , Mice , Mitogens , Peptide Fragments/immunologyABSTRACT
The study of the radiobiological and cytogenetic aspects of induced semi-sterility for application in the genetic control of the onion fly Hylemya antiqua (Meigen) has been continued. Doses of 1.5 krad of X-rays or 0.25 krad of fast neutrons were applied to males and 1.0 krad of X-rays or 0.25 krad of fast neutrons to seven day-old females. On the basis of semi-sterility (between 60% and 30% egg hatch) in backcrosses to normal flies, eleven strains were suspected of carrying a chromosomal rearrangement. Seven had a reciprocal translocation and two from a 1.5 krad X-ray treatment showed complex rearrangements. In two strains no rearrangements were found. Two homozygous translocations are described. Combining data of earlier experiments with the new results we concluded that the irradiation of males with low doses, 0.5 krad of X-rays or 0.25 krad of fast neutrons, is suitable for the induction of chromosomal rearrangements. Strains carrying rearrangements from such low dose treatments will be further used for genetic control experiments on the onion fly.
ABSTRACT
In the preliminary stages of a study into the use of translocations for genetic control of the onion flyHylemya antiqua (Meigen), irradiations were carried out in order to obtain chromosomal rearrangements. Several irradiation experiments, with X-rays or fast neutrons, were carried out on pupae and adults of both sexes at substerilizing doses below 3.0 krad, to establish a favourable way of induction.Because no visible markers are available for the genetic screening of induced rearrangements, and the reciprocal translocations or inversions in demand express themselves in the heterozygous condition by reduced fertility, a total of 234 F1 individuals of both sexes were checked for reduced fertility. 50 F1 individuals were suspected of carrying a translocation or inversion when they produced an egg hatch of between 30 and 60% (semi-sterility).This category was passed for cytogenetic analysis. In the progeny of 25 suspect F1 individuals, 9 different rearrangements were established, of which 7 were translocations. This means a yield of 4% for all the tested F1 After a discussion of the normal karyotype, some of the observed rearrangements are described.Irradiation of males with 1.0 krad of X-rays is advised for the production of semi-sterile stocks carrying translocations. Fast neutrons were not found to be better than X-rays. At doses higher than 1.0 krad complex rearrangements and/or fragments were observed.A translocation homozygote could be isolated in the case of an X-autosomal translocation, and this stock will be used for further genetic control purposes.
