Molecular biological studies: (1) Nematode major sperm protein. (2) Vibrio ribosomal protein L18. (3) Rhodococcus IGTS8 ribosomal RNA. (4) PCR technology

Part 1. Nucleotide sequences of the Major Sperm Protein genes from 21 nematode species were surveyed. The results from this study have provided information about the genomic changes of the highly conserved msp gene during nematode evolution. Analysis of the msp sequence data set is presented. An example is given for the utilization of msp sequences as nematode genetic markers.;Part 2. A strategy suggested by comparative genomic studies was used to amplify the entire Vibrio proteolyticus (Vp) gene for ribosomal protein L18. This approach should allow rapid characterization of L18 from large numbers of bacteria. Both Vp L18 and Escherichia coli (Ec) L18 were overproduced and purified by His-tag affinity chromatography. The purified L18 fusion proteins, were both found to bind to either the Vp 5S or Ec 5S rRNAs in vitro. Vp L18 protein was also shown to incorporate into Ec ribosomes in vivo. This His-tag strategy likely will have general applicability for the study of ribosomal proteins in vitro and in vivo.;Part 3. One of five rRNA gene clusters from Rhodococcus IGTS8 was cloned and the nucleotide sequence determined. Phylogenetic analysis using the 16S rRNA gene suggested this Rhodococcus is a strain or a close relative of an erythropolis species. Complete nucleotide sequence analysis is presented.;Part 4. A mini-review of PCR technology provides an introduction to the description of a method for entrapping PCR reagents in a gel matrix. Studies performed to determine its potential as an effective reactant packaging and delivery system are summarized.