Development and validation of a FACS-based lipoprotein localization screen in the Lyme disease spirochete Borrelia burgdorferi

Table 1 Oligonucleotides used in this study

Number^a	Name	Target/Purpose	Sequence (5' to 3')^b
1	Bsamut-fwd	Introduction of silent mutation in OspA L10 codon yielding Bsa I site	GGGAATAGGTCT C ATATTAGCCTTAATAGC
2	Bsamut-rev	Introduction of silent mutation in OspA L10 codon yielding Bsa I site	TGCTATTAAGGCTAATATG AGACC TATTCC
3	Bstmut-fwd	Introduction of silent mutation in mRFP1 V15R16 codons yielding Bst BI site	TGCGCTTCAAGGT T CG A A TGGAGGGCTCCG
4	Bstmut-rev	Introduction of silent mutation in mRFP1 V15R16 codons yielding Bst BI site	GGAGCCCTCCAT T CG A A CCTTGAAGCGCATGAAC
5	Rmut-oligo	Random mutagenesis oligo	TATTTATTGGGAATAGGTCTC ATATTAGCCTTAATAGCATGTAAGCAAAATGCCTCCTCCNNKNNK GTCATCAAGGAGTTCATGCGCTTCAAGGTTCGAA TGGAGGGCTCCGTG
6	Rmut-rev	Generation of double-stranded DNA from Rmut-oligo	CACGGAGCCCTCCATTCGAA CC
7	Mutscreen-fwd	Amplification of mutated ospA:mrfp1 region from PflaB	ATGCTATTGCTATTTGCGTTTC
8	Mutscreen-rev	Amplification of mutated ospA:mrfp1 region from ospA	ATGGTCTTCTTCTGCATTAC
9	Mutscreen-seq	Sequencing of amplified ospA:mrfp1 region from PflaB	AAAGGATTTGCCAAAGTCAG

ISSN: 1471-2180