Construction Of PiggyBac Transposon Transgene Vector And Studies On Transgenic Methods Of Bombyx Mori

Bombyx mori is an important economical insect. At present,researchers have made it become bioreactor and expressed many kinds ofproteins. Especially, a lot of the silk proteins are specifically andhighly produced in the silk glands. However it still has manydisadvantages, including effect induction,integration,expression ofexogenous gene and steady heredity of transgenic individual, which needto be further researched. Firstly, Transposon expression vector andhelper plasmid were constructed successfully in this thesis. Cytoplasmicactin gene promoter (named A3) from B. mori was fused with recombinedtransposase of piggyBac and a helper plasmid(pARTR1.2)was obtained.Then, the promoter fragment A3 was fused with green fluorescent proteingene (gfp), the fused fragment containing A3 promoter and gfp gene wasinserted into a cloning site between two synthetic transposon piggyBacterminal inverted repeat, constructed a silkworm transgenic vector(pZAGFP) based on piggyBac. The transposon vector has been designeda multiply clone site between the terminal inverted repeat, which becomesmore convenient to be inserted exogenous gene of interest. Secondly, thefibroin light chain cDNA was cloned from the total RNA of Bombyx mori...