TY - JOUR AU - Neuhold, Jana AU - Radakovics, Katharina AU - Lehner, Anita AU - Weissmann, Florian AU - Garcia, Maria Queralt AU - Romero, Mari Carmen AU - Berrow, Nicholas S. AU - Stolt-Bergner, Peggy PY - 2020 DA - 2020/05/12 TI - GoldenBac: a simple, highly efficient, and widely applicable system for construction of multi-gene expression vectors for use with the baculovirus expression vector system JO - BMC Biotechnology SP - 26 VL - 20 IS - 1 AB - Recombinant protein production and purification of large protein complexes in eukaryotes requires efficient methods to generate multi-gene expression constructs, where each individual gene is under the control of its own promoter and terminator. Current methods are based either on serial rounds of combination of several vectors containing loxP sites via the Cre-lox technology, or on multiple rounds of gene combination via PCR or other methods. These methods are multi-step, have lower efficiencies than single gene cloning, and may require laborious processes to verify that all genes of interest are present in the final product. Here, we describe a rapid and simple Golden Gate-based system for the generation of multi-gene expression constructs compatible with baculovirus expression vector systems (BEVS) using either Tn7 transposition or KO1629-based homologous recombination, which we refer to as “GoldenBac”. SN - 1472-6750 UR - https://doi.org/10.1186/s12896-020-00616-z DO - 10.1186/s12896-020-00616-z ID - Neuhold2020 ER -