Constitutive expression of transgenes encoding derivatives of the synthetic antimicrobial peptide BP100: impact on rice host plant fitness

Table 3 Oligonucleotides used in this work

Oligonucleotide code	Sequence	Length
*recursive PCR*
PR1a.1_for	5’ ATA Ggg atc cGA GGC CAC CAT GGG CTT CGT CCT CTT CTC CCA ACT CCC ATC CTT CCT CCT CGT C TC CAC CCT CCT CCT GT 3’	80
BP100.1_ rev	5’ GTA CTT GAG GAT CTT CTT GAA GAG CTT C TT GGC GCG GCA GGA GTG GGA GAT CAC GAG GA A CAG GAG GAG GGT GGA 3’	78
BP100.2_ for	5’ GAA GCT CTT CAA GAA GAT CCT CAA GTA CCT CGC CGG CCC AGC CAA GAA GCT CTT CAA GAA GAT CCT CAA GTA CCT CGC C 3’	82
BP100.3KDEL_ rev	5’ TAT Agg atc cAT TAT CAG AGC TCG TCC TTG AGG TAC TTG AGG ATC TTC TTG AAG AGC TTC TTG GCT GGG CCG GCG AGG TAC TTG AGG 3’	87
*Cloning steps*
PR1a_for	5’ ATA Ggg atc cGA GGC CAC CAT 3’	21
KDELbam_rev	5’ TAT Agg atc cAT TAT CAG AGC TCG TC 3’	26
*Southern blot probe*
SouthUBI_for	5’ ACA TGT GAT GTG GGT TTA CTG ATG 3’	24
SouthBP_rev	5’ GAG GTA CTT GAG GAT CTT CTT GAA G 3’	25
*qPCR*
bp100der_for	5 ’ TCC TCG TGA TCT CCC ACT CCT G 3’	22
bp100der_rev	5’ CGG ATC CAT TAT CAG AGC TCG T 3’	22
hgr_for	5’ CGA AAT TGC CGT CAA CCA AGC 3’	21
hgr_rev	5’ CTG GAG CGA GGC GAT GTT C 3’	19

Underlined sequences indicate overlaps between two oligonucleotides used in recursive PCR, in italics is the pr1a signal sequence, the sequence encoding the KDEL retention signal is indicated in bold, and the BamHI and KpnI restriction sites introduced for cloning purposes are shown in lower case. Length of oligonucleotides is indicated in nt.

ISSN: 1471-2229