By Christopher D. Richardson

Baculovirus Expression Protocols deals either commercial and university-based researchers a accomplished comilation of the newest baculovirus strategies in addition to step by step directions and time-saving recommendations. The contributors-leading specialists within the field-present the various expression plasmids at present in use, consultant the reader during the strategy of producing and choosing recombinant virus, and describe particular examples of recombinant protein construction and purification. The emphasis is on replacement and easier screening thoughts for the choice of recombinant baculovirus. The publication additionally surveys a few of the insect phone traces at the moment appropriate with the baculovirus process. Highlights contain creation of recombinant virus utilizing linearized DNA and vectors that comprise a b-galactosidase indicator and a whole checklist of expression vectors at the moment to be had. special descriptions for the scale-up of protein construction utilizing spinner flasks, bioreactors, and bug larvae also are integrated. Baculovirus Expression Protocols bargains either commercial and university-based researchers a very good number of reproducible, step by step laboratory protocols. it is going to instantly turn into fundamental for an individual operating with baculoviruses and their program within the expression of recombinant proteins in insect cells.

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940 t AGG TTT Baculovirus Transfer Vectors TRANSLATED -360 -350 GAA -290 t AAA TGG CGA -240 ATT CCC AAC ATG -270 TCC GW -230 -220 -170 ATT -150 -100 -80 -40 -30 * PacI* -20 * TTA AAA TAC TAT ACT GTA AAT 20 * * 30 * TTT ACA ATC ATG TCA AAG CCT AAC GTT TTG ACG CAA ATT TTA GAC GCC M S K P N V L T Q I L D A Pi0 protein..... A -50 * 1 -14c * ACT TAC AAC AAG GGG GAC TAT ATT ATC AAA TCA TTT GTA TAT TAA * l AAA ACG CCA AAC GCG TTG GAG TCT 'XT -110 -10 TAC ATT TTA -190 l t -60 * t -200 * -160 l -120 -250 * -210 TAC AAA GAT TCA GAA ATA CGC ATC -70 * -260 * GTT TAC GGG ACT GTG CAA TTG CCG TAC GAT AAA * NBiI * ATT ATG CAT TTG AGG ATG CCG GGA CCT TTA A-A IllRNA.....

It consists of a copy of the polyhedrin gene promoter inserted in tandem with the p10 gene promoter. The transcripts initiating at the p10 promoter are terminated by SV40 signals inserted between the two promoters. The transcripts initiated at the polyhedrin promoter are terminated by the plO- gene signals. The vector also has the facility to produce single-stranded DNA in bacterial cells. Similar vectors (pAcUW42 and pAcUW43) with polylinkers adjacent to the p 10 promoter are also available from PharMingen.

E. (1991) Polyhedrin initiator codon altered to AUU yields unexpected fusion protein from a baculovirus vector. Biotechniques 11,378-383. 26. Sanchez-Martinez, D. and Pellett, P. D. (1991) Expression of HIV-l and HIV-2 glycoprotein G in insect cells by using a novel baculovirus expression vector. Virology 182,229-238. 27. Thiem, S. M. and Miller, L. K. (1990) Differential gene expression mediated by late, very late and hybrid baculovirus promoters. Gene 91,87-94. Baculovirus Transfer Vectors 61 28.

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