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R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata

Journal Article:

Abstract

Site-directed mutagenesis of the photosynthetic apparatus (PSA) genes in Rhodopseudomonas capsulata is presented utilizing a transposon Tn7 mutagenized R-prime. The R-prime, pRPS404, bears most of the genes necessary for the differentiation of the photosynthetic apparatus. Mutagenesis of the R-prime with Tn7 in Escherichia coli, conjugation into R. capsulata, and homologous recombination with the wild-type alleles efficiently generates photosynthetic apparatus lesions. Wild-type alleles are lost spontaneously and the Tn7-induced lesions are revealed by subsequent intramolecular recombination between IS21 insertion elements that bracket the prime sequences in direct repeat. The molecular nature of the intermediates involved in the transposition, recombination and deletion have been investigated by Southern hybridization analysis. The spontaneous loss of wild-type alleles after homologous recombination with the chromosome may be of general use to other prokaryotic site-directed transposon mutagenesis schemes. The IS21-mediated deletion of the prime DNA is dependent on the RecA protein in E. coli, generating the parental R-factor bearing one IS21 element. A genetic-physical map exists for a portion of the prime photosynthetic apparatus DNA. When Tn7 is inserted into a bacteriochlorophyll gene in the R-prime and then crossed into R. capsulata, mutants are produced that accumulate a bacteriochlorophyll precursor, which is in excellent agreement with the  More>>
Authors:
Youvan, D C; [1]  Elder, J T; Sandlin, D E; Zsebo, K; Alder, D P; Panopoulos, N J; Marrs, B L; Hearst, J E
  1. Univ. of California, Berkeley
Publication Date:
Jan 01, 1982
Product Type:
Journal Article
Reference Number:
EDB-84-066585
Resource Relation:
Journal Name: J. Mol. Biol.; (United Kingdom); Journal Volume: 162
Subject:
59 BASIC BIOLOGICAL SCIENCES; ESCHERICHIA COLI; GENETICS; PHOTOSYNTHESIS; GENES; PSEUDOMONAS; GENETIC MAPPING; MUTANTS; PHENOTYPE; PHOTOSYNTHETIC MEMBRANES; PLASMIDS; BACTERIA; BIOLOGY; CELL CONSTITUENTS; CHEMICAL REACTIONS; MAPPING; MEMBRANES; MICROORGANISMS; PHOTOCHEMICAL REACTIONS; SYNTHESIS; 550400* - Genetics; 550700 - Microbiology; 550200 - Biochemistry
OSTI ID:
5128643
Country of Origin:
United Kingdom
Language:
English
Contract Number:
W-7405-ENG-48
Other Identifying Numbers:
Journal ID: CODEN: JMOBA
Submitting Site:
HEDB
Size:
Pages: 17-41
Announcement Date:

Journal Article:

Citation Formats

Youvan, D C, Elder, J T, Sandlin, D E, Zsebo, K, Alder, D P, Panopoulos, N J, Marrs, B L, and Hearst, J E. R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata. United Kingdom: N. p., 1982. Web.
Youvan, D C, Elder, J T, Sandlin, D E, Zsebo, K, Alder, D P, Panopoulos, N J, Marrs, B L, & Hearst, J E. R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata. United Kingdom.
Youvan, D C, Elder, J T, Sandlin, D E, Zsebo, K, Alder, D P, Panopoulos, N J, Marrs, B L, and Hearst, J E. 1982. "R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata." United Kingdom.
@misc{etde_5128643,
title = {R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata}
author = {Youvan, D C, Elder, J T, Sandlin, D E, Zsebo, K, Alder, D P, Panopoulos, N J, Marrs, B L, and Hearst, J E}
abstractNote = {Site-directed mutagenesis of the photosynthetic apparatus (PSA) genes in Rhodopseudomonas capsulata is presented utilizing a transposon Tn7 mutagenized R-prime. The R-prime, pRPS404, bears most of the genes necessary for the differentiation of the photosynthetic apparatus. Mutagenesis of the R-prime with Tn7 in Escherichia coli, conjugation into R. capsulata, and homologous recombination with the wild-type alleles efficiently generates photosynthetic apparatus lesions. Wild-type alleles are lost spontaneously and the Tn7-induced lesions are revealed by subsequent intramolecular recombination between IS21 insertion elements that bracket the prime sequences in direct repeat. The molecular nature of the intermediates involved in the transposition, recombination and deletion have been investigated by Southern hybridization analysis. The spontaneous loss of wild-type alleles after homologous recombination with the chromosome may be of general use to other prokaryotic site-directed transposon mutagenesis schemes. The IS21-mediated deletion of the prime DNA is dependent on the RecA protein in E. coli, generating the parental R-factor bearing one IS21 element. A genetic-physical map exists for a portion of the prime photosynthetic apparatus DNA. When Tn7 is inserted into a bacteriochlorophyll gene in the R-prime and then crossed into R. capsulata, mutants are produced that accumulate a bacteriochlorophyll precursor, which is in excellent agreement with the existing genetic-physical map. This corroborates the mutagenesis scheme.}
journal = {J. Mol. Biol.; (United Kingdom)}
volume = {162}
journal type = {AC}
place = {United Kingdom}
year = {1982}
month = {Jan}
}