title: Use of bacterial DHFR-II fusion proteins to elicit specific antibodies.
abstract: Plasmids containing the coding region of the type II dihydrofolate reductase (DHFR) specified by R388 have been used to alter the amino acid (aa) sequence at the C-terminus of this protein. These plasmids have a unique cloning site in the C-terminal portion of the 78-aa coding region. Insertions of DNA fragments into this site produced plasmids that code for proteins with 6- to 80-aa extensions. The vectors were constructed to terminate translation in all three phases beyond the position of insertion of foreign DNA. Random DNA fragments from the major sperm protein (MSP) gene of Caenorhabditis elegans produced by DNase I cleavage were inserted into these vectors. Cell extracts from colonies containing MSP sequences were examined by gel electrophoresis and immunoblotting. One of the hybrid DHFR-MSP proteins was isolated and antibody was prepared to it. This antibody preparation reacted with MSP in immunoblots of purified MSP and whole cell extracts of the worm. A rapid purification procedure for the DHFR is presented.
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies; D000936:Antigen-Antibody Complex; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005822:Genetic Vectors; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics* / Q000276:immunology
chemical_list: D000906:Antibodies; D000936:Antigen-Antibody Complex; D013762:Tetrahydrofolate Dehydrogenase; D004262:DNA Restriction Enzymes