Generation and in vitro assembly evaluation of a site-specific deletion in Escherichia coli small subunit ribosomal RNA Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/3x816q63k

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  • Ribosomes are intricate macromolecular complexes which are a major element of the protein biosynthetic machinery in all life forms. In Escherichia coli they contain about 50 distinct proteins and 3 ribosomal RNAs. The small 30S ribosomal subunit in E. coli incorporates 21 proteins and a 16S rRNA. The 16S rRNA associated with this subunit was the focus of the investigations described in this dissertation. In order to explore the functional properties of this rRNA an in vitro procedure was developed to generate site-specific internal deletions in the RNA. The C-1400 region of the 16S rRNA was selected for manipulation because the sequence in this zone of the molecule has been shown to be intrinsically universal in all sequenced small subunit rRNAs. Through the use of synthetic DNA, RNase H, and RNA ligase, a four-nucleotide deletion between positions 1400 and 1405 was constructed. The manipulated RNA was tested for competency in in vitro ribosome reconstitution experiments and yielded particles which manifest a sedimentation coefficient comparable to normal 30S subunits. Therefore, this portion of the conserved sequence did not emerge to be a ribosome assembly imperative and must fulfill an essential function during translation.
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  • File scanned at 300 ppi (Monochrome, 8-bit Grayscale) using ScandAll PRO 1.8.1 on a Fi-6770A in PDF format. CVista PdfCompressor 5.0 was used for pdf compression and textual OCR.
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  • description.provenance : Approved for entry into archive by Patricia Black(patricia.black@oregonstate.edu) on 2013-06-10T18:56:29Z (GMT) No. of bitstreams: 1 YooYoungSook1987.pdf: 1051350 bytes, checksum: 076af038f2e0f8e60f98cecca49c6de9 (MD5)
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