2002FEMS Microbiology LettersRequires access

BanAI a new isoschizomer of the type II restriction endonucleaseHaeIII discovered in aBacillus anthracisisolate from Amazon Basin

Jocelei Maria Chies, Ana Christina de Oliveira Dias, Hélio Mauro Moreira Maia, Spartaco Astolfi‐Filho

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Abstract

Bacillus anthracis was isolated and identified from a bacterial collection of samples from the Amazon river bank. Type II restriction endonuclease activity was detected in this prokaryote, the enzyme was purified, the molecular mass of the native protein estimated by gel filtration, and optima pH, temperature and salt requirements were determined. Quality control assays showed complete absence of 'non-specific nucleases'. Restriction cleavage analysis and DNA sequencing of restriction fragments allowed unequivocal demonstration of 5'-GG downward arrow CC-3' as the recognition sequence. This enzyme was named BanAI and is therefore an isoschizomer of the prototype restriction endonuclease HaeIII. This is the first report of a type II restriction endonuclease identified, purified from a natural isolate of B. anthracis.

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Bacillus anthracis was isolated and identified from a bacterial collection of samples from the Amazon river bank. Type II restriction endonuclease activity was detected in this prokaryote, the enzyme was purified, the molecular mass of the native protein estimated by gel filtration, and optima pH, temperature and salt requirements were determined. Quality control assays showed complete absence of 'non-specific nucleases'. Restriction cleavage analysis and DNA sequencing of restriction fragments allowed unequivocal demonstration of 5'-GG downward arrow CC-3' as the recognition sequence. This enzyme was named BanAI and is therefore an isoschizomer of the prototype restriction endonuclease HaeIII. This is the first report of a type II restriction endonuclease identified, purified from a natural isolate of B. anthracis.

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Available abstract

Bacillus anthracis was isolated and identified from a bacterial collection of samples from the Amazon river bank. Type II restriction endonuclease activity was detected in this prokaryote, the enzyme was purified, the molecular mass of the native protein estimated by gel filtration, and optima pH, temperature and salt requirements were determined. Quality control assays showed complete absence of 'non-specific nucleases'. Restriction cleavage analysis and DNA sequencing of restriction fragments allowed unequivocal demonstration of 5'-GG downward arrow CC-3' as the recognition sequence. This enzyme was named BanAI and is therefore an isoschizomer of the prototype restriction endonuclease HaeIII. This is the first report of a type II restriction endonuclease identified, purified from a natural isolate of B. anthracis.

Key concepts: Isoschizomer, Restriction enzyme, HaeIII, Bacillus anthracis, Biology, Restriction fragment, Recognition sequence, Molecular biology

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BanAI a new isoschizomer of the type II restriction endonucleaseHaeIII discovered in aBacillus anthracisisolate from Amazon Basin — Research Paper | ScholarLens