scholarly journals Characterization of Bacillus subtilis Mutants Temperature-sensitive in the Synthesis of Ribonucleic Acid

Microbiology ◽  
1980 ◽  
Vol 120 (2) ◽  
pp. 539-543
Author(s):  
G. MAZZA ◽  
P. PLEVANI
Keyword(s):  
Bacillus Subtilis ◽  
Ribonucleic Acid ◽  
Temperature Sensitive

scholarly journals Bacillus subtilis mutant temperature sensitive in the synthesis of ribonucleic acid.

1976 ◽  
Vol 127 (2) ◽  
pp. 679-690 ◽  
Author(s):  
S Riva ◽  
G Villani ◽  
G Mastromei ◽  
G Mazza
Keyword(s):  
Bacillus Subtilis ◽  
Ribonucleic Acid ◽  
Temperature Sensitive

scholarly journals In Vivo Random Mutagenesis of Bacillus subtilis by Use of TnYLB-1, a mariner-Based Transposon

2006 ◽  
Vol 72 (1) ◽  
pp. 327-333 ◽  
Author(s):  
Yoann Le Breton ◽  
Nrusingh Prasad Mohapatra ◽  
W. G. Haldenwang
Keyword(s):  
Bacillus Subtilis ◽  
Southern Hybridization ◽  
Random Mutagenesis ◽  
Temperature Sensitive ◽  
Transposase Gene ◽  
Gram Positive Bacteria ◽  
Insertion Sites ◽  
Random Transposition

ABSTRACT This report describes the construction and characterization of a mariner-based transposon system designed to be used in Bacillus subtilis, but potentially applicable to other gram-positive bacteria. Two pUC19-derived plasmids were created that contain the mariner-Himar1 transposase gene, modified for expression in B. subtilis, under the control of either σA- or σB-dependent promoters. Both plasmids also contain a transposable element (TnYLB-1) consisting of a Kanr cassette bracketed by the Himar1-recognized inverse terminal repeats, as well as the temperature-sensitive replicon and Ermr gene of pE194ts. TnYLB-1 transposes into the B. subtilis chromosome with high frequency (10−2) from either plasmid. Southern hybridization analyses of 15 transposants and sequence analyses of the insertion sites of 10 of these are consistent with random transposition, requiring only a “TA” dinucleotide as the essential target in the recipient DNA. Two hundred transposants screened for sporulation proficiency and auxotrophy yielded five Spo− clones, three with insertions in known sporulation genes (kinA, spoVT, and yqfD) and two in genes (ybaN and yubB) with unknown functions. Two auxotrophic mutants were identified among the 200 transposants, one with an insertion in lysA and another in a gene (yjzB) whose function is unknown.

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