Table 1.
Primers used for cloning of full-size or deletion mutants of SLAMF1, TRAM, TLR4, and Rab11 FIP2.
ConstructFor/RevPrimer sequence (5′–3′)Restriction enzymePCR product mapped on nucleotide sequence
SLAMF1 (NM_003037.3)     
pcDNA3.1 and pLVX-EF1α-IRES-ZsGreen1 For TTCGAATTCTGATGGGATCCCAAGGGGCTCC EcoRI 367–1,374 
 Rev GCAGCGGCCGCTCAGCTCTCTGGAAGTGTCA NotI  
C-terminal DYKDDDDK For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,371 
 Rev GGTACCTCGAGAGCTCTCTGGAAGTGTCACAC XhoI  
pGEX-2TK For TATGGATCCCAGTTGAGAAGAAGAGGTAAAACG BamHI 1,141–1,374 
 Rev ATAGAATTCTCAGCTCTCTGGAAGTGTCAC EcoRI  
SLAMF1 (NM_003037.3) deletion mutants to C-terminal DYKDDDDK     
All deletion mutants For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI N/a 
1–265 aa Rev GGTACCTCGAGATTTACCTCTTCTTCTCAACTGTAG XhoI 367–1,161 
1–326 aa Rev GTACCTCGAGAGACTGTGATGGAATTTGTTTCCTG XhoI 367–1,344 
1–330 aa Rev GTACCTCGAGACACACTAGCATAGACTGTGATG XhoI 367–1,356 
SLAMF1 (NM_003037.3) deletion mutant to pcDNA3.1     
1–265 aa For TTCGAATTCTGATGGGATCCCAAGGGGCTCC EcoRI 367–1,161 
 Rev TTAAGCGGCCGCTCATTTACCTCTTCTTCTCAACTG NotI  
SLAMF1 (NM_003037.3) mutants with amino acid substitutions to C-terminal DYKDDDDK     
SLAMF1 Y281F For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,225 
 Rev GTTTCTGGACTTGGGCAAAGATCGTAAGGC   
 For GCCTTACGATCTTTGCCCAAGTCCAGAAAC  1,196–1,371 
 Rev GGTACCTCGAGATTTACCTCTTCTTCTCAACTGTAG XhoI  
SLAMF1 Y327F For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,371 
 Rev GGTACCTCGAGAGCTCTCTGGAAGTGTCACACTAGCAAAGACTGTG XhoI  
SLAMF1 TNSI/PNPT For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,341 
 Rev TGTGGTGGGGTTTGGTTCCTGGACAGACTCTGG   
 For GAACCAAACCCCACCACAGTCTATGCTAGTGTGACACTTC  1,324–1,371a 
 Revb CTTGTCATCGTCGTCCTTG   
TRAM/TICAM-2 (NM_021649.7)     
C-terminal DYKDDDDK For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–1,147 
 Rev TTAACTCGAGCGGCAATAAATTGTCTTTGTACC XhoI  
TRAM deletion mutants to C-terminal DYKDDDDK     
1–68 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–646 
 Rev TTAACTCGAGCCATCTCTTCCACGCTCTGAGC XhoI  
1–79 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–679 
 Rev TTACCTCGAGAGAGGAACACCTCTTCTTCAGC XhoI  
1–90 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–712 
 Rev TTACCTCGAGATGTGTCATCTTCTGCATGCAATATC XhoI  
1–100 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–742 
 Rev TTACCTCGAGATAGCAGATTCTGGACTCTGAGG XhoI  
1–120 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–802 
 Rev TTACCTCGAGACTGTCTGCCACATGGCATCTC XhoI  
68–235 aa For CATGAATTCATGTTTGAAGAAGAAGCTGAA EcoRI 644–1,147 
 Rev TTAACTCGAGCGGCAATAAATTGTCTTTGTACC XhoI  
68–176 aa For CATGAATTCATGTTTGAAGAAGAAGCTGAA EcoRI 644–970 
 Rev TTAACTCGAGCCAGGGGCCGCATGGGTATAACAG XhoI  
158–235 aa For CATGAATTCATGAACTCCGTTAACAGGCAGC EcoRI 914–1,147 
 Rev TTAACTCGAGCGGCAATAAATTGTCTTTGTACC XhoI  
TLR4 (NM_138554.4)     
C-terminal DYKDDDDK For CGGTCGACCGAGATCTCATGATGTCTGCCTCGCGCCTGG N/a 299–2,815 
 Rev CTTGTAGTCGCCGGTACCGATAGATGTTGCTTCCTGCCAATTG N/a  
Mus Musculus TRAM/Ticam-2 (NM_173394.3) to EGFP-N1     
1–232 aa For ACTAAGCTTATGGGTGTTGGGAAGTCTAAAC HindIII 477–1,175 
 Rev ATATGGATCCCGGGCAATGAACTGTTTCTGCGAC BamHI  
M. Musculus Slamf1 (NM_013730.4) from pDisplay vector to C-terminal DYKDDDDK     
30–343 aa For TTGGAATTCGGCTTGGGGATATCCACCATGG EcoRI 183–1,127 
 Rev TATACCTCGAGAGCTCTCTGGCAGTGTCACACTG XhoI  
Rab11 FIP2 (NM_014904.2) and deletion mutants to N-terminal DYKDDDDK     
All constructs For GCCCGAATTCGGCTGTCCGAGCAAGCCCAAAAG EcoRI  
1–512 aa Rev ATAGCGGCCGCTCATTAACTGTTAGAGAATTTGCCAGC NotI 446–1,980 
1–327 aa Rev ATAGCGGCCGCTCATTCGCTGCTTTCTTCAAATGG NotI 446–1,429 
1–290 aa Rev ATAGCGGCCGCTTACACAATGCTGTCAGGTTGG NotI 446–1,310 
1–254 aa Rev ATAGCGGCCGCTTATCCGAGAAGATGTGTTTGACC NotI 446–1,199 
1–192 aa Rev ATAGCGGCCGCTTAGTGAGTACTTGGAATGATTGC NotI 446–1,016 
Rab11a (NM_004663.4) QL and SN mutants to pECFP-C1     
1–216 aa For ATCAGTCGACATGGGCACCCGCGACGAC SalI 128–145 
 Rev TTAAGGATCCTTATATGTTCTGACAGCACTG BamHI 758–774 
ConstructFor/RevPrimer sequence (5′–3′)Restriction enzymePCR product mapped on nucleotide sequence
SLAMF1 (NM_003037.3)     
pcDNA3.1 and pLVX-EF1α-IRES-ZsGreen1 For TTCGAATTCTGATGGGATCCCAAGGGGCTCC EcoRI 367–1,374 
 Rev GCAGCGGCCGCTCAGCTCTCTGGAAGTGTCA NotI  
C-terminal DYKDDDDK For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,371 
 Rev GGTACCTCGAGAGCTCTCTGGAAGTGTCACAC XhoI  
pGEX-2TK For TATGGATCCCAGTTGAGAAGAAGAGGTAAAACG BamHI 1,141–1,374 
 Rev ATAGAATTCTCAGCTCTCTGGAAGTGTCAC EcoRI  
SLAMF1 (NM_003037.3) deletion mutants to C-terminal DYKDDDDK     
All deletion mutants For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI N/a 
1–265 aa Rev GGTACCTCGAGATTTACCTCTTCTTCTCAACTGTAG XhoI 367–1,161 
1–326 aa Rev GTACCTCGAGAGACTGTGATGGAATTTGTTTCCTG XhoI 367–1,344 
1–330 aa Rev GTACCTCGAGACACACTAGCATAGACTGTGATG XhoI 367–1,356 
SLAMF1 (NM_003037.3) deletion mutant to pcDNA3.1     
1–265 aa For TTCGAATTCTGATGGGATCCCAAGGGGCTCC EcoRI 367–1,161 
 Rev TTAAGCGGCCGCTCATTTACCTCTTCTTCTCAACTG NotI  
SLAMF1 (NM_003037.3) mutants with amino acid substitutions to C-terminal DYKDDDDK     
SLAMF1 Y281F For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,225 
 Rev GTTTCTGGACTTGGGCAAAGATCGTAAGGC   
 For GCCTTACGATCTTTGCCCAAGTCCAGAAAC  1,196–1,371 
 Rev GGTACCTCGAGATTTACCTCTTCTTCTCAACTGTAG XhoI  
SLAMF1 Y327F For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,371 
 Rev GGTACCTCGAGAGCTCTCTGGAAGTGTCACACTAGCAAAGACTGTG XhoI  
SLAMF1 TNSI/PNPT For TTAGAATTCATGGATCCCAAGGGGCTCC EcoRI 367–1,341 
 Rev TGTGGTGGGGTTTGGTTCCTGGACAGACTCTGG   
 For GAACCAAACCCCACCACAGTCTATGCTAGTGTGACACTTC  1,324–1,371a 
 Revb CTTGTCATCGTCGTCCTTG   
TRAM/TICAM-2 (NM_021649.7)     
C-terminal DYKDDDDK For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–1,147 
 Rev TTAACTCGAGCGGCAATAAATTGTCTTTGTACC XhoI  
TRAM deletion mutants to C-terminal DYKDDDDK     
1–68 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–646 
 Rev TTAACTCGAGCCATCTCTTCCACGCTCTGAGC XhoI  
1–79 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–679 
 Rev TTACCTCGAGAGAGGAACACCTCTTCTTCAGC XhoI  
1–90 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–712 
 Rev TTACCTCGAGATGTGTCATCTTCTGCATGCAATATC XhoI  
1–100 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–742 
 Rev TTACCTCGAGATAGCAGATTCTGGACTCTGAGG XhoI  
1–120 aa For CATGAATTCATGGGTATCGGGAAGTCTAAA EcoRI 443–802 
 Rev TTACCTCGAGACTGTCTGCCACATGGCATCTC XhoI  
68–235 aa For CATGAATTCATGTTTGAAGAAGAAGCTGAA EcoRI 644–1,147 
 Rev TTAACTCGAGCGGCAATAAATTGTCTTTGTACC XhoI  
68–176 aa For CATGAATTCATGTTTGAAGAAGAAGCTGAA EcoRI 644–970 
 Rev TTAACTCGAGCCAGGGGCCGCATGGGTATAACAG XhoI  
158–235 aa For CATGAATTCATGAACTCCGTTAACAGGCAGC EcoRI 914–1,147 
 Rev TTAACTCGAGCGGCAATAAATTGTCTTTGTACC XhoI  
TLR4 (NM_138554.4)     
C-terminal DYKDDDDK For CGGTCGACCGAGATCTCATGATGTCTGCCTCGCGCCTGG N/a 299–2,815 
 Rev CTTGTAGTCGCCGGTACCGATAGATGTTGCTTCCTGCCAATTG N/a  
Mus Musculus TRAM/Ticam-2 (NM_173394.3) to EGFP-N1     
1–232 aa For ACTAAGCTTATGGGTGTTGGGAAGTCTAAAC HindIII 477–1,175 
 Rev ATATGGATCCCGGGCAATGAACTGTTTCTGCGAC BamHI  
M. Musculus Slamf1 (NM_013730.4) from pDisplay vector to C-terminal DYKDDDDK     
30–343 aa For TTGGAATTCGGCTTGGGGATATCCACCATGG EcoRI 183–1,127 
 Rev TATACCTCGAGAGCTCTCTGGCAGTGTCACACTG XhoI  
Rab11 FIP2 (NM_014904.2) and deletion mutants to N-terminal DYKDDDDK     
All constructs For GCCCGAATTCGGCTGTCCGAGCAAGCCCAAAAG EcoRI  
1–512 aa Rev ATAGCGGCCGCTCATTAACTGTTAGAGAATTTGCCAGC NotI 446–1,980 
1–327 aa Rev ATAGCGGCCGCTCATTCGCTGCTTTCTTCAAATGG NotI 446–1,429 
1–290 aa Rev ATAGCGGCCGCTTACACAATGCTGTCAGGTTGG NotI 446–1,310 
1–254 aa Rev ATAGCGGCCGCTTATCCGAGAAGATGTGTTTGACC NotI 446–1,199 
1–192 aa Rev ATAGCGGCCGCTTAGTGAGTACTTGGAATGATTGC NotI 446–1,016 
Rab11a (NM_004663.4) QL and SN mutants to pECFP-C1     
1–216 aa For ATCAGTCGACATGGGCACCCGCGACGAC SalI 128–145 
 Rev TTAAGGATCCTTATATGTTCTGACAGCACTG BamHI 758–774 

For, forward; QL, Rab11a Q70L mutant; Rev, reverse; SN, Rab11a S25N mutant.

a

Partially on vector sequence.

b

On C-terminal DYKDDDDK vector.

View Large
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal