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Team:Wageningen UR/Journal/week21
From 2012.igem.org
week 21: 17 September - 23 September
Hepatitis B inside modification
- Colony PCR of Hepatitis B core protein behing an IPTG promoter in pSB1C3 backbone with sequencing primers
-> the gel shows fragments of the expected size (about 1kb)
GFP modification
17 September
- ligation of the PCR products from 14 September (mixing the duplo samples together) with pJET and transformation with JM109
18 September
- colony PCR of the transformation from 17 September
CCMV coil addition
Frame shift in coil addition After recieving our samples from the sequencing, we saw that there was a frame shift in the sequence, leading to the faulty translation of the CCMV delta 25 coat protein, with an early stop codon.
| faulty | intended | |
| BP sequence | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAAGATAGC
GGCGTTGAAGGAGAAAATCGCAGCACTAAAAGAAAAGATAG CGGCGTTGAAGGAGCTTGGTGGTGGTTCTGGTGGTGGTGGT TCTGCTGCTGCTGTGTGGTCCAACCTGTTATTGTAGAACCC ATCGCTTCAGGCCAAGGCAAGGCTATTAAAGCATGGACCGG TTACAGCGTATCGAAGTGGACCGCCTCTTGTGCGGCTGCCG AAGCTAAAGTAACCTCGGCTATAACTATCTCTCTCCCTAAT GAGCTATCGTCCGAAAGGAACAAGCAGCTCAAGGTAGGTAG AGTTTTATTATGGCTTGGGTTGCTTCCCAGTGTTAGTGGCA CAGTGAAATCCTGTGTTACAGAGACGCAGACTACTGCTGCT GCCTCCTTTCAGGTGGCATTAGCTGTGGCCGACAACTCGAA AGATGTTGTCGCTGCTATGTACCCCGAGGCGTTTAAGGGTA TAACCCTTGAACAACTCACCGCGGATTTAACGATCTACTTG TACAGCAGTGCGGCTCTCACTGAGGGCGACGTCATCGTGCA TTTGGAGGTTGAGCATGTCAGACCTACGTTTGACGACTCTT TCACTCCGGTGTAT | GTTTCTTCGAATTCGCGGCCGCTTCTAGATGAAGATAGC
GGCGTTGAAGGAGAAAATCGCAGCACTAAAAGAAAAGATAG CGGCGTTGAAGGAGCTTGGTGGTGGTTCTGGTGGTGGTGGT TCTGCTGCTGCTGGTGTGGTCCAACCTGTTATTGTAGAACC CATCGCTTCAGGCCAAGGCAAGGCTATTAAAGCATGGACCG GTTACAGCGTATCGAAGTGGACCGCCTCTTGTGCGGCTGCC GAAGCTAAAGTAACCTCGGCTATAACTATCTCTCTCCCTAA TGAGCTATCGTCCGAAAGGAACAAGCAGCTCAAGGTAGGTA GAGTTTTATTATGGCTTGGGTTGCTTCCCAGTGTTAGTGGC ACAGTGAAATCCTGTGTTACAGAGACGCAGACTACTGCTGC TGCCTCCTTTCAGGTGGCATTAGCTGTGGCCGACAACTCGA AAGATGTTGTCGCTGCTATGTACCCCGAGGCGTTTAAGGGT ATAACCCTTGAACAACTCACCGCGGATTTAACGATCTACTT GTACAGCAGTGCGGCTCTCACTGAGGGCGACGTCATCGTGC ATTTGGAGGTTGAGCATGTCAGACCTACGTTTGACGACTCT TTCACTCCGGTGTAT |
| primer FW1 FL-Delta26 | TGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGCTGC
TGT'GTGGTCCAACCTGTTATTGTAG | TGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGCTGC
TGGTGTGGTCCAACCTGTTATTGTAG |
| translation product (AA) | MKIAALKEKIAALKEKIAALKELGGGSGGGGSAAAVW
SNLLL | MKIAALKEKIAALKEKIAALKELGGGSGGGGSAAAGV
VQPVIVEPIASGQGKAIKAWTGYSVSKWTASCAAAEAKV TSAITISLPNELSSERNKQLKVGRVLLWLGLLPSVSGTV KSCVTETQTTAAASFQVALAVADNSKDVVAAMYPEAFKG ITLEQLTADLTIYLYSSAALTEGDVIVHLEVEHVRPTFD DSFTPVY |
the new primer is ordered to still make this part available for the registry, but submission before the wiki deadline will not be possible anymore. however, we think that iGEM should not be the only reason to deliver bricks, and we intend to deliver all bricks that did not make it just in time, still after the iGEM deadlines.
