Isolation of an asporogenic (spoOA) protective antigen-producing strain of Bacillus anthracis

1999 ◽  
Vol 45 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Patricia L Worsham ◽  
Michele R Sowers
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacillus Anthracis ◽  
Congo Red ◽  
Southern Hybridization ◽  
Protective Antigen ◽  
Essential Gene ◽  
Vaccine Production ◽  
Chain Reaction ◽  
Safe Production ◽  
Polymerase Chain

We found that Congo red agar allows identification of sporulation-deficient Bacillus anthracis. Using Congo red agar, we isolated an asporogenic derivative of the protective antigen-producing strain B. anthracis deltaSterne-1(pPA102). Polymerase chain reaction and Southern hybridization analyses of DNA from the asporogenic mutant revealed that a deletion was present in spoOA, an essential gene for the initiation of sporulation. The deletion also encompassed the spoIVB homologue and a portion of the recN homologue. The avirulent spoOA strain deltaSterne-1(pPA102)CR4 is suitable for the safe production of protective antigen without endospore contamination of the vaccine production facility.Key words: Bacillus anthracis, protective antigen, spoOA, vaccine, Congo red.

Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

LWT ◽  
2015 ◽  
Vol 63 (1) ◽  
pp. 714-719 ◽  
Author(s):  
Sahilah Abd Mutalib ◽  
Nursheila Mustafa Muin ◽  
Aminah Abdullah ◽  
Osman Hassan ◽  
Wan Aida Wan Mustapha ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Southern Hybridization ◽  
Pcr Analysis ◽  
Conventional Pcr ◽  
Chain Reaction ◽  
Gelatin Capsules ◽  
Halal Authentication ◽  
Polymerase Chain

The use of deficiencies to determine essential gene content in the let-56–unc-22 region of Caenorhabditis elegans

Genome ◽  
1993 ◽  
Vol 36 (6) ◽  
pp. 1148-1156 ◽  
Author(s):  
Jacquie E. Schein ◽  
Marco A. Marra ◽  
Guy M. Benian ◽  
Chris Fields ◽  
David L. Baillie
Keyword(s):  
Polymerase Chain Reaction ◽  
Caenorhabditis Elegans ◽  
Genomic Sequence ◽  
Essential Gene ◽  
Chain Reaction ◽  
Radiation Induced ◽  
Practical Tool ◽  
Polymerase Chain ◽  
Selection Of

We have investigated the possibility of using the polymerase chain reaction to detect deletions of coding elements in the unc-22–let-56 interval on chromosome IV in the nematode Caenorhabditis elegans. Our analysis of approximately 13 kb of genomic sequence immediately to the left of the unc-22 gene resulted in the identification of four possible genes. Partial cDNAs have been identified for three of them. To determine whether any of these coding elements are essential for development, we required a method for the induction and selection of mutations in these elements. Our approach was to identify a set of formaldehyde and gamma radiation induced unc-22 mutations that mapped to the unc-22–let-56 region, and then employ polymerase chain reaction methodology to identify deficiencies that affected one or more of the four identified coding elements. Two small deficiencies were identified in this manner. Characterization of these deficiencies shows that there are no coding elements between unc-22 and let-56 (the nearest mutationally identified gene to the left of unc-22), which are required in development under laboratory conditions. We conclude that the polymerase chain reaction is a practical tool for the detection of deletions of coding elements identified in this region, and that characterization of such deficiencies provides a method for assessing whether or not these elements are required for development.Key words: Caenorhabditis elegans, deficiencies, coding elements, unc-22.

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