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Table 1 List of different regulatory elements used to design a set of gag piggyBac vector constructs.

From: Use of the piggyBac transposon to create HIV-1 gag transgenic insect cell lines for continuous VLP production

Construct

Enhancer

Promoter

Intron upstream

Gene

Intron downstream

PolyA

Final pXLBacII construct

pHSP70

 

Drosophila hsp70

 

gag

 

SV40sti/polyA

pXLHSP70Gag

pIE1

hr5

Baculovirus AcMNPV ie1

 

gag

 

(Heliothis) hel2polyA

pXLIE1Gag

pIE1

hr5

Baculovirus AcMNPV ie1

 

Neo

 

hel2polyA

pXLNeo

pIE1-SV

hr5

Baculovirus AcMNPV ie1

 

gag

 

SV40sti/polyA

pXLIE-SVGag

pActin

 

Drosophila actin 5C

 

gag

 

hel2-polyA

pXLActinGag

pActin-SV

 

Drosophila actin 5C

 

gag

 

SV40sti/polyA

pXLActin-SVGag

409-FOR

hr5

Baculovirus AcMNPV ie1

+

gag

 

hel2polyA

pXL409Gag

410-FOR

hr5

Baculovirus AcMNPV ie1

 

gag

+

hel2polyA

pXL410Gag

411-FOR

 

Drosophila actin 5C

+

gag

 

hel2-polyA

pXL411Gag

412-FOR

 

Drosophila actin 5C

 

gag

+

hel2-polyA

pXL412Gag

Hr3ieLuc

hr3

Baculovirus AcMNPV ie1

 

gag

  

pXlHr3ieGag

pBSII-IE1-orf

 

Baculovirus AcMNPV ie1

 

gag

 

pBSII-IE1-orf polyA

pXLBSII-IE1Gag

  1. The promoters tested included Drosophila hsp70, AcMNPV ie1 and Drosophila actin 5C [30, 31] (column 3). The promoter enhancers used were hr5 [31] and hr3 [30] (column 2). Poly-adenylation (polyA) sequences included SV40sti/polyA, hel2polyA [31] and a polyA tail isolated from the pBSII-IE1-orf helper plasmid [14, 15] (column 7). 409-FOR and 410-FOR are pIE1 vectors containing a 133 bp intron [32] (column 4 and 6) upstream or downstream of the gag gene, respectively. 411-FOR and 412-FOR are pActin vectors containing the intron upstream or downstream of the gag gene, respectively. Briefly, different promoters, enhancers, polyA sequences or the intron were removed from the construct described in column 1 and coupled to gag in the transposable pXLBacII vector [14, 15] (column 8).