Some partial sequences were extended to full length using 5' and/or 3' RACE.
Several partial cDNAs of the P450s and UGTs were extended to full length by 5' and/or 3' rapid amplification of cDNA ends (RACE).
The P450 sequences ZfCYP4G47, ZfCYP4G48, ZfCYP4L17, ZfCYP6AE27, ZfCYP6CT1, CYP9A36, ZfCYP9A37, ZfCYP304F2, ZfCYP332A3, ZfCYP333B8, ZfCYP379A2, and ZfCYP379A3 were verified by Race-PCR using the SMART RACE cDNA Amplification Kit from Clontech (cat. 634914), and partial sequences were extended to full length.
This observation was recently extended to full-length LmrA in proteoliposomes [31].
At the highest enzyme concentrations used (32 nM) 72% of control templates were extended to full-length product.
As previously observed in the running start primer extension assays, these 22-mer products were also not extended to full-length products by the action of hRev1 alone.
In fact, this property extends to full-length receptors; some pLGICs in invertebrates are activated by acetylcholine as well as serotonin [ 6].
Since all of the genes encode relatively long proteins, most of the collection comprises gene fragments which cannot yet be extended to full-length.
The M1dG lesion blocked incorporation of the initial nucleotide opposite the lesion, but 14% of templates were extended to full-length product.
At the same time, the detected gene fragments might be extended to full-length genes with further improvement of the genome assembly.
The leading strand is continuously extended to full-length product; consequently, once Pol α has switched to the DNA elongation mode, it does not appear to switch back to the priming mode.
