A polyphosphate kinase 1 (ppk1) mutant of Pseudomonas aeruginosa exhibits multiple ultrastructural and functional defects -- Fraley et al. 104 (9): 3526 Data Supplement - HTML Page - index.htslp -- Proceedings of the National Academy of Sciences

Fraley et al. 10.1073/pnas.0609733104.

Supporting Information

Files in this Data Supplement:

SI Movie 1
SI Table 3

SI Movie 1

Movie 1. Real-time video microscopy of the P. aeruginosa ppk1 mutant (Left) and WT (Right) in rich (LB) medium. Motilities were quantified digitally (see Materials and Methods).

Table 3. PCR primers used in this study to clone P. aeruginosa ppk1 and verify the chromosomal deletion/insertion mutation

Primer	Sequence	Restriction site
Cloning
F2	gcgAAGCTTTCCCTTACCGCCTTCAAACG	HindIII
R2	gcgTCTAGAGCAGAGCCCAAGAGCCTTTC	XbaI
F6	gcgAAGCTTCCCTCGGGAAGATGAATGAATACG	HindIII
R6	gcgGATATCTCAACGTGCGGTAAGCACCGG	EcoRV
Verification
F4	ACAGCGACGAGGCACTTCAC
R4	CATAACCACTACCACAAGCAGGG
F5	AGGGCAACAAGGCCACCTAC
R5	ACCTGATGGAGCACTCCGAG
TF1	CAGGTAGATGACGACCATCAG
TR1	CCAAGTAGCGAAGCGAGCAG

All sequences are given 5' to 3'; endonuclease restriction sites are in bold. The ppk1 ATG start and TGA stop codons in primers F6 and R6, respectively, are underlined. All primers were generated by the GCG program Primer using either the pPPK02F or entire P. aeruginosa PAO1 genome sequences and confirmed for uniqueness against the entire genome sequence.