TABLE 3.
Oligonucleotides used in this study
| Oligonucleotides a | Sequence (5’ – 3’) | Description b |
|---|---|---|
| oVPL329 | attccttggacttcatttactgggtttaac | Rev for screening pJP_Leptin_tag |
| oVPL659 | tgccccgttagttgaagaag | Fwd for pSIP411 |
| oVPL660 | attctgctcccgcccttatg | Rev for pSIP411 |
| oVPL1221 | gcttgaaacgttcaattgaaatggca | Screening oligonucleotide for pJP_Leptin_tag |
| oVPL1326 | ccagttggtaacaatgccatgt | Sequencing oligonucleotide for pSIP_Leptin_tag_EFTu_LgBiT |
| oVPL1366 | taaagcaattactgatattgctg | Fwd for EFTu amplification |
| oVPL1447 | cgaattaatagaaaaacattagtcaaatac | Fwd for pJP backbone |
| oVPL1448 | taatgaaaacctcctgataatttacaag | Rev for pJP backbone |
| oVPL2115 | aacacaagcattacgtaaactca | Rev for IL-22 amplification |
| oVPL2238 | atggttccaattcaaaaagttcaagatg | Sequencing oligonucleotide for pJP_Leptin_tag |
| oVPL2239 | atgttaccagttaatactcgttgtaaat | Fwd for IL-22 amplification |
| oVPL2351 | taatctcgctttgattgttctatcg | Sequencing oligonucleotide for pJP_Leptin_tag |
| oVPL4252 | tgatctttgaaccaaaattagaaaacc | Fwd for pJP backbone |
| oVPL4254 | tatttaaaaaaattagttgatctttgaaccaaaattagaaaacc | Fwd starting at stop codon of leptin with 17 bases of tag sequence |
| oVPL4255 | aacgccaaccactaacacattctggactaacatctaattgttg | Rev upstream of stop codon leptin with 16 bases of tag sequence |
| oVPL4307 | gctttcgatagaacaatcaaagcgagattattaagagttgatagtaacacgaaataacat | Bridging oligonucleotide to fuse LgBiT and pJP backbone |
| oVPL4308 | accaacgaaatcctcaagagtaaataccattaatgaaaacctcctgataatttacaagta | Bridging oligonucleotide to fuse LgBiT and EFTu |
| oVPL4309 | gtatttgactaatgtttttctattaattcgtaaggaagataaatcccataagggcgggag | Bridging oligonucleotide to fuse pJP backbone and EFTu |
| oVPL4310 | taaggaagataaatcccataaggg | Fwd for pJP backbone |
| oVPL4312 | tcatcaagtgttatatagcggtc | Rev for LgBiT screening |
| oVPL4336 | cattgagaagattgccgaaa | Fwd for LgBiT |
| oVPL4337 | atttgatccgctgacaatcc | Sequencing oligonucleotide for LgBiT and EFTu |
| oVPL4339 | cccgtctaaggaattgtcagat | Sequencing oligonucleotide for pSIP_Leptin_tag_EFTu_LgBiT |
| oVPL4418 | gttagtggttggcgtttatttaa | Fwd for pJP_Leptin_tag backbone |
| oVPL4493 | atgattaattataaacaattacaattac | Fwd for IFN-β amplification |
| oVPL4493 | atgattaattataaacaattacaattac | Screening oligonucleotide for IFN-β |
| oVPL4494 | attttgaaaattacgagttaaacgacg | Rev for IFN-β amplification |
| oVPL4538 | caaccatatatgcaagaagt | Sequencing oligonucleotide for IL-22 |
| oVPL4538 | caaccatatatgcaagaagt | Screening and sequencing oligonucleotide for IL-22 |
| oVPL4539 | cgtgatttattacatttattagc | Sequencing oligonucleotide for Leptin |
| oVPL4540 | ttggcgtgttcaacgttatt | Sequencing oligonucleotide for IFN-β |
| oVPL4540 | ttggcgtgttcaacgttatt | Sequencing oligonucleotide for oligo for IFN-β |
| oVPL4543 | ttgattacctccttatcatca | Rev for pSIP_Leptin_tag |
| oVPL4544 | tctagactcgaggaattcggt | Fwd for pSIP_Leptin_tag |
| oVPL4848 | aaacgccaaccactaaccattaatgaaaacctcctgataatt | Fwd starting at stop codon of IFN-β with 17 bases of tag sequence |
| oVPL4849 | atttaaaaaaattagtattaattataaacaattacaattacaa | Rev upstream of stop codon IFN-β with 16 bases of tag sequence |
| Recombinant DNA | ||
| gVPL71 | atggtatttactcttgaggatttcgttggtgactgggaacagactgccgctta taatttagatcaagtattagaacaaggcggcgtgtcttcattgctccagaac ttggcagtttcagttactcctattcaacggattgttcggagtggggaaaatgc gctgaagattgatattcatgttattattccgtacgaaggattgtcagcggatc aaatggcacaaatagaagaagtttttaaagtggtatatccggtcgatgatc atcatttcaaagttattttaccttacgggaccttagtaattgatggtgtgactcc taatatgttaaactattttggacgtccttatgaaggaattgctgtttttgatggaa agaagattacagttacaggaactttgtggaatggaaataagattatcgatg agagattaattactccagatggaagtatgttatttcgtgttactatcaactcttaa |
Codon optimized LgBiT |
a
oVPL, van Pijkeren Laboratory primer identification number.
b
Fwd, forward oligonucleotide; Rev, reverse oligonucleotide.