Table 2.
Primers used in this study
| Primer | Sequencea | Sense | Application |
|---|---|---|---|
| 8a-F1 | 5′-CGGGATCCGCCACCATGAAACTTCTC-3′ | + | pXJ-8a and pXJ-8ab |
| 8a-F2b | 5′-CGGGATCCACCATGGGAATGAAACTTCTC-3′ | + | pXJ-8a-myc and pXJ-8ab-myc |
| 8a-F3 | 5′-CGGGATCCACCATGGGAATATGCACTGT-3′ | + | pGex4T1-8aΔN15 |
| 8a-R1 | 5′-CCGCTCGAGCTAGTGTTGTACC-3′ | − | pXJ-8a and pGex4T1-8aΔN15 |
| 8a-R2 | 5′-CCGCTCGAGTTGTGTTGTACC-3′ | − | pXJ-8a-myc |
| 8b-F1 | 5′-CGGGATCCGCCACCATGTGCTTGAAG-3′ | + | pXJ-8b |
| 8b-F2b | 5′-CGGGATCCACCATGGGAATGTGCTTGAAG-3′ | + | pXJ-8b-myc |
| 8b-F3 | 5′-CGGGATCCACCATGGGAGTTTTACCTTT-3′ | + | pGex4T1-8bΔN26 |
| 8b-R1 | 5′-CCGCTCGAGTTAATTTGTTCGT-3′ | − | pXJ-8b and pXJ-8ab and pGex4T1-8bΔN26 |
| 8b-R2 | 5′-CCGCTCGAGCCATTTGTTCGTTTATT-3′ | − | pXJ-8b-myc and pXJ-8ab-myc |
| 8ab-1 | 5′-GTTGGTACCCAGTAGGACAAGGATCTTC-3′ | − | Construction of 8ab |
| 8ab-2 | 5′-GGTTACCAACCTGAATGGAATATAAGG-3′ | + | Construction of 8ab |
| 8ab-3 | 5′-TCCATTCAGGTTGGTACCCAG-3′ | − | Construction of 8ab |
| 8ab-4 | 5′-AATGGAATATAAGGTACAACAC-3′ | + | Construction of 8ab |
| 8ab-5 | 5′-CCTTATATTCCATTCAGG-3′ | − | Construction of 8ab |
a
Restriction sites introduced into primers are shown in bold face.
b
Six additional base pairs (ATGGGA), which encodes for two additional amino acids (methionine and glycine), were added to the 5′ end of 8a-myc, 8b-myc and 8ab-myc to give a Kozak consensus ribosome binding site for more efficient translation initiation.