Table 1.
Vector | Fusion tag | Parent vector/ antibiotic resistance | Promoters/baculoviral recombination sites | Forward primer extension | Reverse primer extension |
---|---|---|---|---|---|
pOPINE | C-terminal … KHHHHHH | pTriEx2/ampicillin | T7lacO, CMV enhancer and β-actin promoter, p10 promoter/ lef-2 and 1629 baculo elements. | AGGAGATATACCATG† | GTGATGGTGATGTTT† |
pOPINF* | N-terminal MAHHHHHHSSGLEVL FQGP … | pTriEx2/ampicillin | T7lacO, CMV enhancer and ββ-actin promoter, p10 promoter/ lef-2 and 1629 baculo elements. | AAGTTCTGTTTCAGGGCCCG‡ | ATGGTCTAGAAAGCTTTA‡ |
pOPING | N-terminal MGILPSPGMPALLSLV SLLSVLLMGCVAET G … cleavable secretion leader and C-terminal … KHHHHHH | pTriEx2/ampicillin | (T7lacO-not used), CMV enhancer and β-actin promoter, p10 promoter/lef-2 and 1629 baculo elements. | GCGTAGCTGAAACCGGC | GTGATGGTGATGTTT |
pOPINJ* | N-terminal MAHHHHHHSSG-GST- LEVLFQÞGP … | pTriEx2/ampicillin | T7lacO, CMV enhancer and β-actin promoter, p10 promoter/lef-2 and 1629 baculo elements. | AAGTTCTGTTTCAGGGCCCG‡ | ATGGTCTAGAAAGCTTTA‡ |
pOPINM* | N-terminal MAHHHHHHSSG-MBP- LEVLFQGP … | pTriEx2 ampicillin | T7lacO, CMV enhancer and β-actin promoter, p10 promoter/lef-2 and 1629 baculo elements. | AAGTTCTGTTTCAGGGCCCG‡ | ATGGTCTAGAAAGCTTTA‡ |
-represents the point of cleavage by 3C protease or signal peptidase (as appropriate). Vectors marked ‡ use the same primer extensions, enabling the same PCR product to be cloned into all marked vectors. Underlined sequences represent methionine initiation or stop codons (as appropriate) and may be excluded from the gene-specific primers.