Expression of cauliflower mosaic virus gene I in insect cells using a novel polyhedrin-based baculovirus expression vector.

摘要: An improved polyhedrin-based baculovirus expression vector was constructed to expedite distinguishing infections by putative recombinants from wild-type (wt) baculovirus. The utilizes the Escherichia coli beta-galactosidase gene (lacZ) as a genetic marker for positive recombination between wt Autographa californica nuclear polyhedrosis virus and transfer vector. gene/expression cassette so that lacZ deleted polyhedrin were transcribed in opposite orientations, both terminating simian 40 DNA fragment which acts bidirectional terminator. In vector, is Drosophila melanogaster heat-shock promoter (hsp 70), constitutively expressed baculovirus-infected Spodoptera frugiperda (Sf) cells, thereby making site of available insertion foreign genes under control promoter. Recombinant baculoviruses are readily selected plaque assays development blue colour upon addition X-Gal. selection renders retrieval less dependent on high frequency DNA. usefulness this new illustrated expressing I cauliflower mosaic virus, encodes protein Mr 46,000. Expression at same level cells infected with conventional Gene formed large hollow fibre-like structures cytoplasm Sf cells. This first plant be insect recombinant