Open AccessCloning, DNA sequence, and expression of the Rhodobacter sphaeroides light-harvesting B800-850-alpha and B800-850-beta genes
Author(s) -
Patricia J. Kiley,
Samuel Kaplan
Publication year - 1987
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.169.7.3268-3275.1987
Subject(s) - biology , operon , rhodobacter sphaeroides , microbiology and biotechnology , gene , molecular cloning , plasmid , nucleic acid sequence , sequence analysis , gene expression , genetics , bacteria , escherichia coli
Two deoxyoligonucleotide probes were synthesized in accordance with the available amino acid sequence of the B800-850-beta polypeptide from Rhodobacter sphaeroides and were used to isolate a 2.6-kilobase PstI fragment from R. sphaeroides 2.4.1 chromosomal DNA. Identification of the B800-850-beta and B800-850-alpha structural genes, pucB and pucA, was confirmed by DNA sequencing. Northern (RNA) blot analysis, using restriction endonuclease fragments from the cloned genes as probes, revealed a single puc-operon-specific, highly stable transcript of approximately 640 bases present in photosynthetically grown cells. In vitro transcription-translation analysis of the puc operon revealed that the maximum synthesis of the puc operon gene products was achieved when the entire 2.6-kilobase PstI fragment was used as the template, although a 537-base-pair XmaIII fragment was sufficient to direct the synthesis of pucB and pucA fusion product.