Method for introducing specific and unmarked mutations into the chromosome of Streptococcus pneumoniae
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Last Update: 2021-03-06
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Source: Internet
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Author: User
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This work describes a procedure for the generation of site-specific mutations into the chromosome of
Streptococcus pneumoniae
that does not involve the use of an antibiotic resistance marker. A linear fragment of transforming deoxyribonucleic acid (
DNA
) is constructed by polymerase chain reaction (
PCR
) (gene splicing by overlap extension) and used to transform competent cells of
S. pneumoniae
. Selection of transformants is performed by PCR, and typically, 1% of the transformed cells show the expected mutation. By this protocol it is possible to change a single base pair into the pneumococcal genome, as well as obtaining in-frame deletions and insertions.
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