Active center studies of bacterial luciferases by nucleotide sequence analysis and site-directed mutagenesis /
| Main Author: | |
|---|---|
| Other Authors: | , , , |
| Format: | Thesis Book |
| Language: | English |
| Published: |
1989.
|
| Subjects: | |
| Online Access: | Link to OAKTrust copy |
| Abstract: | The nucleotide sequences of the luxA and luxB genes encoding α and β subunits of bacterial luciferases from Vibrio fischeri ATCC 7744 and Photobacterium leiognathi PL741 have been determined. Alignment and comparison of the encoded amino acid sequences of luciferase subunits from V. harveyi, V. fischeri and P. leiognathi showed that 56% of the amino acid residues of the α subunits and 41% of the amino acid residues of the β subunits of the enzymes from the three species are identical. Furthermore, the amino acid sequences of the α and β subunits of luciferases are similar: 13% of the amino acid residues of the α and β subunits of luciferases from the three species are identical, i.e. they were identical at 13% of the comparative positions for all six subunits, confirming that the luciferases are homologous and the α and β subunits of luciferases have arisen from a common ancestor via gene duplication. By cloning of the luxA and luxB genes from two different species into a common vector, it was possible to form hybrid luciferases; the expression of bioluminescence from hybrid luciferases in vivo supplied evidence supporting conservation of the subunit interface. Using this approach, it was possible to form hybrid luciferases that do not form in vitro upon refolding from urea.. |
|---|---|
| Item Description: | Typescript (photocopy). Vita. "Major subject: Biochemistry." |
| Physical Description: | xiv, 118 leaves : illustrations ; 29 cm |
| Bibliography: | Includes bibliographical references. |