Document Detail


Amplified expression of ribulose bisphosphate carboxylase/oxygenase in pBR322-transformants of Anacystis nidulans.
MedLine Citation:
PMID:  2644909     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Prior research suggested that the genes for large (L) and small (S) subunits of ribulose bisphosphate carboxylase/oxygenase (RuBisCO) are amplified in ampicillin-resistant pBR322-transformants of Anacystis nidulans 6301. We now report that chromosomal DNA from either untransformed or transformed A. nidulans cells hybridizes with nick-translated [32P]-pBR322 at moderately high stringency. Moreover, nick-translated [32-P]-pCS75, which is a pUC9 derivative containing a PstI insert with L and S subunit genes (for RuBisCO) from A. nidulans, hybridizes at very high stringency with restriction fragments from chromosomal DNA of untransformed and transformed cells as does the 32P-labeled PstI fragment itself. The hybridization patterns suggest the creation of two EcoRI sites in the transformant chromosome by recombination. In pBR322-transformants the RuBisCO activity is elevated 6- to 12-fold in comparison with that of untransformed cells. In spite of the difference in RuBisCO activity, pBR322-transformants grow in the presence of ampicillin at a similar initial rate to that for wild-type cells. Growth characteristics and RuBisCO content during culture in the presence or absence of ampicillin suggest that pBR322-transformants of A. nidulans 6301 are stable. The data also collectively suggest that a given plasmid in the transformed population replicates via a pathway involving recombination between the plasmid and the chromosome.
Authors:
H Daniell; J A Torres-Ruiz; A Inamdar; B A McFadden
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Archives of microbiology     Volume:  151     ISSN:  0302-8933     ISO Abbreviation:  Arch. Microbiol.     Publication Date:  1989  
Date Detail:
Created Date:  1989-03-14     Completed Date:  1989-03-14     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0410427     Medline TA:  Arch Microbiol     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  59-64     Citation Subset:  IM    
Affiliation:
Biochemistry/Biophysics Program, Washington State University, Pullman 99164-4660.
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MeSH Terms
Descriptor/Qualifier:
Ampicillin Resistance / genetics
Cyanobacteria
Gene Amplification
Gene Expression Regulation
Genes, Bacterial*
Plasmids
Ribulose-Bisphosphate Carboxylase / genetics*
Transformation, Genetic
Grant Support
ID/Acronym/Agency:
GM-19,972/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
EC 4.1.1.39/Ribulose-Bisphosphate Carboxylase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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