By combining the HAEC and BAC (Bacterial Artificial Chromosome) systems, we designed and constructed the second generation HAEC vector, which can shuttle large DNA inserts i.e. at least up to 260 kb between bacteria and human cells. Such a system is being used to develop large insert libraries, whose clones can be directly transferred into mammalian cells for functional analysis. Since the HAEC DNAs are easy to isolate from mammalian cells, we can transfer the BAC- HAEC clone DNA from mammalian cells into bacterial cells. This facilitates the procedures of cloning disease genes.
Currently, we are using this vector to conduct several functional cloning studies including the construction of a total human genomic HAEC library and several region-specific HAEC libraries by subcloning YACs or BAC/PAC/P1 clones.