SYNTHESIS OF OLIGONUCLEOTIDES

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Synthesis of oligonucleotides by our laboratory in collaborative efforts with several laboratories within the Division of Bacterial Products has proven invaluable to the following research programs: A PCR approach to clone and express a potential surface antigen from Mycoplasma pneumonia, and to modify existing cloning vectors. Development of a method to allow for direct cloning of PCR generated products, and as templates for DNA sequencing in Mycoplasma. Completion of the nucleotide sequence analysis of mycobacterial genes encoding several immunoreactive antigens. Isolation of the pneumococcal pneumolysin gene from two types of genomic DNA, to produce trunicated point mutations of the pneumolynsin gene, and for conjugation of a single polysaccharide. Continued research in the production of selective mutation and amplification of bacterial toxin genes for use in examining the effect of mutation on toxin function. Primers were also utilized in the development of PCT protocols to rapidly detect non tuberculosos mycobacterial infections. Research in our laboratory is being conducted in the use of cloned overlapping strnads, to develop an in vitro transcription probe for allergenic screening of apple to determine and isolate the allergenic component. Studies are also continuing to include the use of oligonucleotides as tools for the creation of an epitope library.