Both G∶C and A∶T mutations are detected in Jurkat-AID cells.

<p>G∶C and A∶T focused mutations were monitored using <i>p</i>mutEGFP-TAG182, <i>p</i>mutEGFP-TAG52 and <i>p</i>mutEGFP-TAA52 SHM vectors. The experiment was repeated four times for TAG-182 and TAG-52 codons and three times for the TAA-52 codon. The average value of these independent experiments is represented. The percentage of fluorescent cells was calculated relative to transfection efficiency which was monitored using a wild-type EGFP vector. Twenty hours after transfection, 8.1% of Jurkat-AID cells reverted the TAG-182 codon, 27.08% the TAG-52 codon and 13.22% the TAA codon. Less than 1% of Jurkat cells reverted TAG-182 and TAA-52 codons and TAG-52 reverted in 3.97% of the transfected cells.</p

Mutations can be detected very rapidly using SHM vectors.

<p>Jurkat and Jurkat-AID cells were transfected with the <i>p</i>mutEGFP-TAG182 SHM vector. Fluorescent cells were detected 3 and 24 hours after transfection. Three hours after transfection, 0.23% of fluorescent Jurkat-AID cells are observed and 1.68% had reverted 24 hours after transfection.</p

Reversion status of plasmids rescued from fluorescent colonies

<p>Reversion status of plasmids rescued from fluorescent colonies</p

Data_supplement_Murray_Pathgeography

Data underpinning analyses presented Boxe

AID is transcribed in the Jurkat-AID and not in the control Jurkat cell line.

<p>AID transcription was monitored with RT-PCR in a control cell line, Jurkat, and in a Jurkat-AID cell line stably transfected with AID. A Burkitt lymphoma cell line Ramos constitutively expressing AID was used as a positive control for AID expression. G3PDH was used as an internal control. The bands corresponding to AID (380 bp) and G3PDH (1000 bp) are shown with black arrows.</p

The <i>p</i>mutEGFP-TAG182 SHM vector was tested in Jurkat and Jurkat-AID cell lines.

<p>Twenty hours after transfection cells were analyzed on a FACS Scan. Cells that have reverted the stop codon of one or more copies of the vector appear fluorescent. The percentage of fluorescent cells relative to transfection efficiency was monitored using a wild-type EGFP vector. The experiment was repeated 4 times. An average of 8.1% of transfected Jurkat-AID cells reverted the stop codon, compared to 0.29% of non-transfected Jurkat cells.</p

H4 - H4.nex

Aligned sequences for H4 avian subtyp

H2 - H2.nex

Aligned sequences for H1 avian subtyp

H12

Aligned sequences for H12 avian subtyp

H3 - H3.nex

Aligned sequences for H3 avian subtyp