Rapid targeted gene disruption in Bacillus anthracis

BMC Biotechnology

Table 2 Primers used for retargeting

Primer name	Primer sequence	Pairings changed
IS605-435\|436 s	AAAAAAGCTTATAATTATCCTTAAAATACAGTGTAGTGCGCCCAGATAGGGTG	IBS1/2
IS605-435\|436 s	CAGATTGTACAAATGTGGTGATAACAGATAAGTCAGTGTAGATAACTTACCTTTCTTTGT	EBS1/δ
IS605-435\|436 s	TGAACGCAAGTTTCTAATTTCGGTTTATTTCCGATAGAGGAAAGTGTCT	EBS2
BAS4553-343\|344 s	AAAAAAGCTTATAATTATCCTTAGATGCCCGTACGGTGCGCCCAGATAGGGTG	IBS1/2
BAS4553-343\|344 s	CAGATTGTACAAATGTGGTGATAACAGATAAGTCCGTACGGCTAACTTACCTTTCTTTGT	EBS1/δ
BAS4553-343\|344 s	TGAACGCAAGTTTCTAATTTCGATTGCATCTCGATAGAGGAAAGTGTCT	EBS2
BAS4597-1794\|1795 s	AAAAAAGCTTATAATTATCCTTACGAAACGAGAAAGTGCGCCCAGATAGGGTG	IBS1/2
BAS4597-1794\|1795 s	CAGATTGTACAAATGTGGTGATAACAGATAAGTCGAGAAAGATAACTTACCTTTCTTTGT	EBS1/δ
BAS4597-1794\|1795 s	TGAACGCAAGTTTCTAATTTCGGTTTTTCGTCGATAGAGGAAAGTGTCT	EBS2
EBS-Universal	AACCGAAATTAGAAACTTGCGTTCA	None

The primers shown were used in PCR to generate a HindIII BsrG1 fragment that was exchanged into the Targetron vectors and retarget the intron to the genomic location indicated in the primer name. The exact nucleotide the intron is designed to insert at is designated by a | at the genomic location indicated above the primer sequences.

ISSN: 1472-6750