Table 2 Primers for generating PLCε and RhoA mutants
From: RhoA allosterically activates phospholipase Cε via its EF hands
Primer name | Sequence 5′ to 3′ |
|---|---|
RhoAG14V Forward | TGTTGGTGATGTAGCCTGTGGAA |
RhoAG14V Reverse | ATCACCAGTTTCTTCCGG |
EF-C Forward | GAAGGCCCAACCTTGGCA |
EF-C Reverse | CATCTTGACCAGGATCCG |
EF3-C Forward | GCTGCTGCAAGCATTGTG |
EF3-C Reverse | CATCTTGACCAGGATCCGCCC |
PLCΔY Forward | AGAACTCCTAAATGCTATC |
PLCΔY Reverse | TGAATTTAGAGTTGACAGG |
W1051E Reverse | GTGCTCTCTCTCCTACCACCAAACAACTCC |
F1187E Forward | CCAGAGCGAAATGGTCTCAGACAGTAACATGAG |
F1187E Reverse | ACCATTTCGCTCTGGAAGCCTTTCATTCCTC |
V1189K Forward | TTCATGGAATCAGACAGTAACATGAGTTTCATTG |
V1189K Forward | GTCTGATTCCATGAAGCTCTGGAAGCC |
I1279E Forward | GAGGCAGGAATCTGACGCCATTGCTGCTG |
I1279E Reverse | TCAGATTCCTGCCTCTGGTTGTCAGAAAGC |
A1282K Forward | ATCTGACAAAATTGCTGCTGCAAGCATTGTGA |
A1282K Reverse | GCAATTTTGTCAGATATCTGCCTCTGGTTGTCA |
I1283E Forward | TGACGCCGAAGCTGCTGCAAGCATTGTGAC |
I1283E Reverse | GCAGCTTCGGCGTCAGATATCTGCCTCTGG |
A1286K Forward | TGCTGCTAAAAGCATTGTGACTAATGGCACTGG |
A1286K Reverse | ATGCTTTTAGCAGCAATGGCGTCAGATATCTGC |
I1295E Forward | CACTGGGGAAGAAAGCACGTCCCTGGGC |
I1295E Reverse | CTTTCTTCCCCAGTGCCATTAGTCACAATGC |
Δ1275–1289 Forward | TTTCTGACACTAATGGCACTGGGATTGAAAGCA |
Δ1275–1289 Reverse | CATTAGTGTCAGAAAGCTGCTGCTTACTCTTC |
Δ1287–1298 Forward | CTGCTGCATCCCTGGGCATATTTGGGGTC |
Δ1287–1298 Reverse | CCAGGGATGCAGCAGCAATGGCGTC |
Δ1275–1298 Forward | TTTCTGACTCCCTGGGCATATTTGGGGT |
Δ1275–1298 Reverse | CCAGGGAGTCAGAAAGCTGCTGCTTACTCTTC |
T977E Forward | GCTTCAGGAAACCGACAATAGATTACTGCACTTCG |
T977E Reverse | TCGGTTTCCTGAAGCCCATAGAGCAGCG |
F1012E Forward | ACGGAAGGAACCTGACCAAAGACAACAGTGG |
F1012E Reverse | TCAGGTTCCTTCCGTATCTTTCTCACAGC |
F1203E Forward | CGAGCTGGAAAAATCATTCAGCATAAGGAGCCGC |
F1203E Reverse | GATTTTTCCAGCTCGACGAATTCAATGAAAC |
F1206E Forward | CAAATCAGAAAGCATAAGGAGCCGCAAG |
F1206E Reverse | ATGCTTTCTGATTTGAACAGCTCGACG |
R1209E Forward | CAGCATAGAGAGCCGCAAGGACTTGAAGG |
R1209E Reverse | CGGCTCTCTATGCTGAATGATTTGAACAGCTCG |
Q1408E Forward | TCTACAGCGAGGTCCTCTTGCAAGGATG |
Q1408E Reverse | GGACCTCGCTGTAGAGCTCCACAGAGG |
L1460E Forward | CTCCGACGAGCCAATCATCATATCCATTGAGAAC |
L1460E Reverse | ATTGGCTCGTCGGAGGTGATGAAGGCAC |
PLCε ΔEF Forward | AAGGCCCAATGGATAAAGACAATTTCGCCTC |
PLCε ΔEF Reverse | TATCCATTGGGCCTTCATACCGACC |
PLCε/β3 EF Forward | linearize: ATGGATAAAGACAATTTCGCCTC PLCβ3 insert: CGGTATGAAGGCCCATTCCTGCGCAAAGCATACACG |
PLCε/β3 EF Reverse | linearize: TGGGCCTTCATACCGACC PLCβ3 insert: ATTGTCTTTATCCATCTCCTCGCCTCCCAGGTAGC |
PLCε/β3 EF1/2 Forward | linearize: ATGGATAAAGACAATTTCGCCTC PLCε insert: ATCTTTGAGCGGTTCGCTGCAAGCATTGTGACTAATGG |
PLCε/β3 EF1/2 Reverse | linearize: GAACCGCTCAAAGATTTCCAAGG PLCε insert: ATTGTCTTTATCCATCAGAAACCTTGCAAACCCT |
PLCε/β3 EF3/4 Forward | linearize: CTGAACAAGCTGTGTCTGCGG PLCε insert: CGGTATGAAGGCCCAACCTTGGCACATGCTGTGG |
PLCε/β3 EF3/4 Reverse | linearize: TGGGCCTTCATACCGACC PLCε insert: ACACAGCTTGTTCAGAGCAATGGCGTCAGATATCTGCC |
PLCβ3/ε EF2 Forward | linearize: CTGCGGCCGGACATTGAC PLCε insert: ATGTTCTCAGCAGACTTCATTGAATTCGTCGAGCTGT |
PLCβ3/ε EF2 Reverse | linearize: GTCTGCTGAGAACATCTTCAGG PLCε insert: AATGTCCGGCCGCAGCACAATGCTTGCAGCAGCA |
PLCβ3/ε E2α’ Forward | linearize: CTGCGGCCGGACATTGAC PLCε insert: CTGAACAAGCTGTGTCTTTCTGACAACCAGAGGCAGA |
PLCβ3/ε E2α’ Reverse | linearize: ACACAGCTTGTTCAGGAACCG PLCε insert: AATGTCCGGCCGCAGTTCAATCCCAGTGCCATTAGTCACA |
