Detail information of the subunit gene library and polygene vector construction service

The biGBac is a combination of baculovirus Bac to Bac expression and Gibson assembly reaction technology. Despite the Bac to Bac part, three plasmids are involved in the Gibson assembly reaction. This page provides an overview of each plasmid as well as the assembly reaction to help customers understand Creative BioMart’s biGBac expression service.

pLIB Vector Level -Construction of the Subunit Gene Library

In order to further reduce the time consumption of the whole construction process, a universal subunit gene library was constructed.

The individual fragment was cloned into the library vector (pLIB) and flanked by a polyhedrin promoter (prom) in front and a transcriptional terminator sequence (term) in behind. This cassette can be called a gene expression cassette (GEC). A lot of GECs together with the pLIB plasmid consist of the subunit gene library. When researchers want to use some certain gene to do a Gibson assembly, they just need to amplify the gene with the by PCR using sets of predefined primer that decorate the DNA fragment ends with the optimized linker sequences α, β, γ, δ, e, and ω.

pBIG1 Vector Level- Vector Structure and Construction Logic

At this stage, desired DNA fragments are amplified from the library vectors and decorated with α, β, γ, δ, e, and ω linker. These linker sequences then guide the assembling of PCR products in the order of α to ω. The newly formed generated polygene cassette (PGC) is digested by Pmel to release from the plasmid with two novel linkers A and B. Similarly, other PGCs treated with PmeI will also be attached with CDEF linkers. Up to five PGCs can be used to generate larger multigene assemblies in pBIG2.

pBIG2 Vector Level- Vector Structure and Construction Logic

PGCs from different pBIG1 constructs with compatible linker sequences are inserted into PmeI-linearized pBIG2 vectors. Again, up to five PGCs from different pBIG1 are combined with pBIG2 to generate the ultimate pBIG2 constructs can be used to transfer its GECs onto a baculoviral genome using Tn7 transposition.

We determine the price according to the complexity of the foreign gene insertion procedure.

Want more gene insertion? Please contact us or turn to the online inquiry to let us know your request, Creative BioMart will always be your powerful research assistance. Also, if you are interested in Creative BioMart's biGBac polygene expression service or have any question, please feel free to contact us.