Team:Freiburg Bioware/testpage

(Difference between revisions)

Revision as of 10:42, 6 September 2010

banner

pAAV-MCS vector (Stratagene): we survey the vector for igEM-restiction sites (RFC-25)
EcoRI 1327; Xbal 1344; are inside the MCS
NgoMIV 2254 are inside the V1 Ori region
function of the Beta-Globin intron ( pAAV MCS Vector): increases the expression rate in Vivo/ Vitro with several mechanisms ( i.a. increased m-RNA accumulation (Media: Freigem10_Nott_et_al_A_quantitative_analysis_of_intron_effects_on_mammalian_gene_expression_2003.pdf?; Media: Freigem10_Haddad_et_al_A_systematic_study_of_the_function_of_the_h-beta-globin_introns_on_the_expression_2009.pdf?)
Thymidin Kinase : we received another modified thymídin kinase-sequence from Amor, to do: check for iGEM restriction sites and develop a cloning strategie

Investigators: Adrian, Chris W., Hanna, Bea

tomorrow we will obtain the vector sequence for the thymidinkinase (WT) from Amor
call at Stratagene: #240071 AAV-Helper Free System, 1412 € (netto), available until next week ; we asked for a discount (Bea got Infos per E-mail)
next step (tomorrow, 10 am): modification of the multiple cloning site

Investiators: Adrian, Chris W., Hanna, Kerstin, Anissa, (Kristian, Tobias, Sven)

thymidin kinase from Amor: there is a PstI restriction site within the sequence (instead of this kinase we will probably utilize the TK30 + SR39 mutant -> we have to find their sequences )
analyze of the secundary struckture of the ITR's (pAAV MCS of Stratagene), to decide if we can perform a point mutation to delete the PstI-restriction site : http://www.molbiotech.uni-freiburg.de/iGEM/wiki2010/images/8/82/ITRright.pdf http://www.molbiotech.uni-freiburg.de/iGEM/wiki2010/images/4/48/ITRleft_ohnePstI.pdf http://www.molbiotech.uni-freiburg.de/iGEM/wiki2010/images/e/ef/Freiburg_10ITRleft.pdf http://www.molbiotech.uni-freiburg.de/iGEM/wiki2010/images/b/b5/Freiburg_10ITRright_AAV2.pdf
sequence alignement of the beta-globin of the pAAV MCS Plasmid with human beta-globin: http://www.molbiotech.uni-freiburg.de/iGEM/wiki2010/images/8/83/Freiburg10_Nucleotide_alignment_221_extraction.pdf
To do: gather informations about the mutant thymidinkinase (TK30 + SR39)

we won't use this kinase...

@@ Line 122: / Line 122: @@
 <div class="box_right">
 </html>
-====2. Labday 13.04.2010====
+===2. Labday 13.04.2010===
-===<p style="font-size:15px; background-color:#66bbFF;"><b>The Biobricks Foundation: RFC</b></p>===
+====<p style="font-size:15px; background-color:#66bbFF;"><b>The Biobricks Foundation: RFC</b></p>====
 <p><b>Investigators: Chris W., Bea, Kira, Hanna, Julian, Anna, Jessica (Tobias,Sven, Kristian)</b></p><br>
@@ Line 182: / Line 182: @@
 <br>
-====4. Labday 22.04.2010====
+===4. Labday 22.04.2010===
-===<p style="font-size:15px; background-color:#66bbFF;"><b>Theoretical cloning</b></p>===
+====<p style="font-size:15px; background-color:#66bbFF;"><b>Theoretical cloning</b></p>====
 <p><b>Investigators: Adrian, Chris W., Hanna, Bea</b></p><br>
@@ Line 194: / Line 194: @@
 <br>
-====5. Labday 23.04.2010====
+===5. Labday 23.04.2010===
-===<p style="font-size:15px; background-color:#66bbFF;"><b>Theoretical cloning</b></p>===
+====<p style="font-size:15px; background-color:#66bbFF;"><b>Theoretical cloning</b></p>====