携改良型HBVpre-SI基因表达载体的构建及相关-研究.pdf

formedintoDH5a cellsof the competentprepared，in wassendto extractionand recombinant purification，theplasmid sequence， then AH109 lithiumacetatemethod theuseoftransformedcells yeast by the transformedon sizeof ceilingSD／Trpscreening．Pick(2～3)mm coloniesafter culture，the extraction，analyzedby overnight yeastprotein andthe ofasodium sulfate conventionalmethodsexpressionproduct dodecyl Westemblot analysis． gelelectrophoresis(SDS)and polyacrylamide strains adr’S 1 Results：The HBV HBVpre—Sgenefragment，PCR amplified revealedthat were electrophoresisanalysis ampli- 1．5％agarosegel products fled ofabout3 withthe expectedfragment，and fragments 57bp，consistent PCR HBV By amplification nothingspecificamplificationphenomenon 1 showedthat iscorrect．Byoverlap pre．Sgenesequenceanalysis sequence extension two PCR ofDNAobtain— independentamplification method，using