Genes encoding for fusion proteins were assembled as a multicomponent cloning cassette from annealed oligonucleotides (IDT DNA) containing these elements (in order): BglII or NheI or BamHI — Secretion Signal/FLAG tag — HindIII — (αDTX or DTX-K or CONK1) — KpnI — Linker — LOV2-Jα(404–546) — NotI — PDGF-R-mCherry — XbaI or EcoRI. PDGF-R-mcherry was derived from pFU-MVIIA-PC (Addgene)10 (link). See Table 1 for sequence details. This cassette was inserted into the mammalian expression vector pcDNA3.1(+) (Invitrogen) using NheI / Xba restriction sites or a lentiviral vector containing the CAMKII promoter35 (link),36 (link) using BamHI / EcoRI sites. The respective genes coding for rat Kv1.2 (Kv1.2), rat Kv1.1 (Kv1.1) or Shaker were amplified from Kv1.2-pBluescript, Shaker-pBluescript (gifts from Roderick Mackinnon), or BacMam Kv1.1 (Invitrogen) and inserted into pcDNA3.1(+) or pEGFP-N3 using BamHI / EcoRI or NheI/EcoRI, respectively. Both αDTX-lumitoxin and Kv1.2 cassettes were also inserted into the bidirectional expression vector pBI-CMV1 (Clontech) using BglII / XbaI and BamHI / NotI sites respectively, to drive expression from the same plasmid.
Pfu mviia pc
Manufactured by Addgene
The PFU-MVIIA-PC is a laboratory instrument designed for the purification of proteins. It utilizes a specific column resin to selectively capture and concentrate target proteins from complex biological samples. The core function of this product is to enable efficient protein purification for downstream analysis and research applications.
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Engineered Optogenetic Fusion Proteins
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Engineered Optogenetic Fusion Proteins
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Genes encoding for fusion proteins were assembled as a multicomponent cloning cassette from annealed oligonucleotides (IDT DNA) containing these elements (in order): BglII or NheI or BamHI — Secretion Signal/FLAG tag — HindIII — (αDTX or DTX-K or CONK1) — KpnI — Linker — LOV2-Jα(404–546) — NotI — PDGF-R-mCherry — XbaI or EcoRI. PDGF-R-mcherry was derived from pFU-MVIIA-PC (Addgene)10 (link). See Table 1 for sequence details. This cassette was inserted into the mammalian expression vector pcDNA3.1(+) (Invitrogen) using NheI / Xba restriction sites or a lentiviral vector containing the CAMKII promoter35 (link),36 (link) using BamHI / EcoRI sites. The respective genes coding for rat Kv1.2 (Kv1.2), rat Kv1.1 (Kv1.1) or Shaker were amplified from Kv1.2-pBluescript, Shaker-pBluescript (gifts from Roderick Mackinnon), or BacMam Kv1.1 (Invitrogen) and inserted into pcDNA3.1(+) or pEGFP-N3 using BamHI / EcoRI or NheI/EcoRI, respectively. Both αDTX-lumitoxin and Kv1.2 cassettes were also inserted into the bidirectional expression vector pBI-CMV1 (Clontech) using BglII / XbaI and BamHI / NotI sites respectively, to drive expression from the same plasmid.
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