A series of T7 promoters described previously [1, 2, 11, 12], see Table 1 for sequences, were placed upstream of the broccoli coding sequence via primer extension. Sense strand primers with promoter sequences,
(Graumann & Premstaller, 2006). The T7 promoter is incorporated into theexpression vectorsuch that it controls the transcription of the heterologous gene; the T7 promoter is only recognized byT7 RNA polymeraseand, thus, the gene coding for T7 RNA polymerase is introduced into thebacterial ...
Cross-linking of promoter DNA to any of the native cysteines would allow the RNA polymerase to survive the high salt challenge, but function with DNA would almost certainly happen in trans, as it should not allow correct positioning of the promoter on the protein to which it is cross-linked...
We cloned the PCR product containing a 28 nt fragment of the mboIIM2ΔA356 gene, including deletion of A356 which generated stop codon TAA, fused with the GFP en- coding gene (35) in a pET24a vector (Supplementary Ta- bles S1 and S2). This allowed us to identify...