Golden Gate cloning enables efficient concatemer construction for biophysical analysis of heterozygous potassium channel variants from patients with epilepsy.

• Tác giả: Alexey R Brazhe, Andrei M Gigolaev, Daria A Iureva, Stanislav V Lagosha, Alexander A Vassilevski, Boris S Zhorov
• Ngôn ngữ: eng
• Ký hiệu phân loại: 338.456 Production efficiency in specific industries and groups of industries
• Thông tin xuất bản: Netherlands : International journal of biological macromolecules , 2025
• Bộ sưu tập: NCBI
• ID: 723055
• Chủ đề: 1. Golden; 2. Gate; 3. Cloning

Tóm tắt

Missense mutations that cause channelopathies usually occur in a heterozygous setting. Functional voltage-gated potassium channels are tetramers of pore-forming α-subunits. When a variant is co-expressed with the wild-type gene, six distinct tetramers can assemble. Conventional techniques cannot be used to study the tetramers individually because they rely on simultaneous expression of separate genes resulting in a mixture of all possible combinations. An established approach is generation of concatemeric constructs, but it is technically challenging and laborious for long cDNA. Here, we chose a missense variant of K