TABLE 1.

Oligonucleotide primers used

Primer	Oligonucleotide sequencea	Annealing temp (°C)b
LppT-NdeI-N	AACATATGAAAAAACCTAATTTTAAAACAT	54
LppT-NotI-C	TCGCGGCCGCTCTTAGCAATTGCTTGTCTTGC	57
LppT-mut1L	GATTTTTGGTTTGAGCAAAGAGA	54
LppT-mut1R	CTCAAACCAAAAATCAAATTTTTG	51
LppT-mut2L	TTCCAATGGGAAGCTGACCAA	56
LppT-mut2R	AGCTTCCCATTGGAATTTATTG	53
RGD-mutL	GCCCGTGGTGAAGTTTAC	51
RGD-mutR	GTAAACTTCACCACGGGC	51
RGD-reverse	TTTTGTAAACGGCATCAACTACGACATC	60
RGD-forward	GTTGATGCCGTTTACAAAACTGTAGTAG	59

^aUnderlined nucleotides represent restriction enzyme sites, and nucleotides presented in boldface indicate site-specific mutations.

^bObtained with the Oligonucleotide Properties Calculator (http://www.basic.northwestern.edu/biotools/oligocalc.html), using the nearest neighbor method and the parameters 300 nM primer and 50 mM salt (Na⁺), not considering nucleotides added to create the restriction enzyme recognition sites.