Characterization of a Novel Group I Intron in Bacillus Bacteriophage FS7

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; R o o t E n t r y F o/ @ C o m p O b j b W o r d D o c u m e n t O b j e c t P o o l o/ @ o/ @ 4 @ 9 : ; = ? @ A B F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; 9520503 Wilson PRIVATE This proposal is for a Career Advancement Award to extend the PIs sabbatical leave from Siena College for one semester; to provide release time from teaching for an additional semester upon return to her home institution; and to provide summer salary support. She wishes to continue a promising line of research initiated during a sabbatical in Dr. David Shub's laboratory at the State University of New York at Albany. During the proposed period of support she also plans to write and submit a research grant on this project to the NSF Research in Undergraduate Institutions program so that she can continue the investigations in with undergraduates. A systematic search for introns will be undertaken to explore the evolutionary relationships between Group I introns in prokaryotic genes. A recently discovered self splicing intron within the thymidylate synthase (TS) gene of B. subtilis bacteriophage ,B22 (thy) shares a number of properties with the previously identified thymidylate synthase (td) Group I intron of bacteriophage T4, as well as with the other known T4 introns (nrdB and sunY). A number of candidate phages of gram(+) bacteria, identical in structure to ~22, will be explored by PCR for introns within their TS genes. Each new intron discovered will be cloned into an in vitro transcription vector to analyze its ability to self splice from primary transcripts, and sequenced to deduce its folded structure, and ascertain its relatedness to the known Group I introns. %%% Although the vast majority of eukaryotic genes contain introns, none has been identified as residing in eubacteria or their viruses until the surprising discovery of a self splicing intron in the thymidylate synthase gene of bacteriophage T4. Others in T4 were quickly discovered as well. All of the intron containing genes in T4 encode proteins involved in the synthesis of deoxyribonucleotides in infected host cells. In the Career Advancement Award, a systematic search for introns in prokaryotic genes will be undertaken. each new intron identified will be analyzed for its ability to self-splice, and its structures and relatedness to other group I introns will be determined. *** ; @ ....()()))()() Z : phoenix W EH o ; S u m m a r y I n f o r m a t i o n ( 8 Oh +' 0 $ H l D h R:\WWUSER\TEMPLATE\NORMAL.DOT marcia steinberg marcia steinberg @ t @ @ @ @ @ J 6 Microsoft Word 6.0 3 e = e " " " " " " " L L L L L d n L C x | | | | | | | # f W T 4 " | | | | | | " " | x | | | | " | " | 6 " " " " | | 5 | 9520503 Wilson PRIVATE This proposal is for a Career Advan