Generation and in Vitro Assembly Evaluation of a Site-specific Deletion in Escherichia Coli Small Subunit Ribosomal RNA PDF Download
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Author: Young Sook Yoo
Publisher:
ISBN:
Category : Escherichia coli
Languages : en
Pages : 168
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Book Description
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.
Author: Young Sook Yoo
Publisher:
ISBN:
Category : Escherichia coli
Languages : en
Pages : 168
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Book Description
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.
Author: Fazilah Abdul-Latif
Publisher:
ISBN:
Category : Escherichia coli
Languages : en
Pages : 132
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Book Description
The ribosome is a central component of the protein synthetic apparatus. It is a macromolecular complex of protein and RNA. Although much progress has been made in understanding the functional role of the proteins in this particle, little is known of the functional role which the RNA plays. Naturally occurring primary structural mutations in rRNAs have not been reported. The work described here focused on developing methodology for generating site-specific deletions in rRNA directly to explore the functional properties of the RNA. The 3'-terminus of E. coli small subunit 16S rRNA was chosen as the site to be deleted. This region of the RNA is believed to be important in the initiation of protein synthesis and could be essential for proper assembly of ribosomes. The deletions were achieved by synthesizing a DNA molecule of 10 bases in length which was complementary to the 3'-end of 16S rRNA. The DNA was hybridized to the RNA and then the RNA component of the hybrid was specifically digested with a combination of RNase H isolated from E. coli and calf thymus. This produced an equal mixture of "16S" RNAs missing either 10 or 8 terminal nucleotides. This RNA was shown to be functional in in vitro reconstitution studies indicating that this zone of the RNA is not essential for ribosome assembly.
Author: Andrew Scheinman
Publisher:
ISBN:
Category : Escherichia coli
Languages : en
Pages : 288
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Book Description
Author:
Publisher:
ISBN:
Category : Dissertation abstracts
Languages : en
Pages : 768
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Book Description
Author:
Publisher:
ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 1252
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Author: Gary Spedding
Publisher: IRL Press
ISBN:
Category : Science
Languages : en
Pages : 352
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Book Description
A practical and self-contained introduction to methods of researching the structure and function of the ribosome in light of the increasing recognition of the potential capability of RNA molecules to act as molecular catalysts. Also describes protein synthesis and cell-free synthesizing systems. Annotation copyrighted by Book News, Inc., Portland, OR
Author:
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages : 1860
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Book Description
Author: Knud H. Nierhaus
Publisher: John Wiley & Sons
ISBN: 9783527306381
Category : Science
Languages : en
Pages : 608
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Book Description
Knud Nierhaus, who has studied the ribosome for more than 30 years, has assembled here the combined efforts of several scientific disciplines into a uniform picture of the largest enzyme complex found in living cells, finally resolving many decades-old questions in molecular biology. In so doing he considers virtually all aspects of ribosome structure and function -- from the molecular mechanism of different ribosomal ribozyme activities to their selective inhibition by antibiotics, from assembly of the core particle to the regulation of ribosome component synthesis. The result is a premier resource for anyone with an interest in ribosomal protein synthesis, whether in the context of molecular biology, biotechnology, pharmacology or molecular medicine.
Author: Patrick Franklin Suthers
Publisher:
ISBN:
Category :
Languages : en
Pages : 188
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Book Description
Author: Boyd Hardesty
Publisher: Springer Science & Business Media
ISBN: 1461248841
Category : Science
Languages : en
Pages : 832
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Book Description
During the past few decades we have witnessed an era of remarkable growth in the field of molecular biology. In 1950 very little was known of the chemical constitution of biological systems, the manner in which information was transmitted from one organism to another, or the extent to which the chemical basis of life is unified. The picture today is dramati cally different. We have an almost bewildering variety of information de tailing many different aspects of life at the molecular level. These great advances have brought with them some breath-taking insights into the molecular mechanisms used by nature for replicating, distributing, and modifying biological information. We have learned a great deal about the chemical and physical nature of the macromolecular nucleic acids and proteins, and the manner in which carbohydrates, lipids, and smaller mole cules work together to provide the molecular setting of living systems. It might be said that these few decades have replaced a near vacuum of information with a very large surplus. It is in the context of this flood of information that this series of mono graphs on molecular biology has been organized. The idea is to bring together in one place, between the covers of one book, a concise assessment of the state of the subject in a well-defined field.
