Manual for the u of
脱口秀笑星黄西
mating-bad Split ubiquitin system
"mbSUS"中秋节的英文
version A
Petr Obrdlik
October 2004
Introduction and general information
amor什么意思
Abbreviations
X = prey peptide/ORF
Y = bait prey/ORF
N = NubG
天使用英语怎么说C = Cub
PLV = protA-LexA-VP16 peptide
HA tag = hemagglutinin epitope tag
Ade = adenine
His = histidine
Trp = tryptophan
Leu = leucine
Ura = uracil
Met = methionine
X-Gal = 5-bromo-4-chloro-3-indol-b-D-galactosida ONPG = o-nitrophenylgalactopyranoside
pant
SC = synthetic complete medium
万达 石景山
SD = synthetic dextro / minimal medium SSDNA = carrier DNA from salmon sperm
mbSUS package contents
You have received the package with mating-bad Split ubiquitin system "mbSUS". This package contains:
yeast strains THY.AP4 and THY.AP5
vectors (resuspended in dH
2O, ca. 50 ng/ul)
1pXNgate21-3HA 2pNXgate33-3HA 3pX-NubWTgate 4pNubWT-Xgate 5pMetYCgate
control constructs (resuspended in dH
2O, ca. 50 ng/ul)
6KAT1-Cub (Arabidopsis K+ channel, Acc. At5g46240)
vaginacontest>魔术贴 英文
7KAT1-N
11N-KAT1-3HA
Restriction analysis:
The maps of the vectors and control constructs are shown in the ction "Maps of vectors and control constructs". The quences of the vectors1-5 in genebank format are in the "Appendix".flight
General information, protocols, maps and quences of the vectors are included in this manual. For further information plea check Obrdlik et
al., 2004 and Ludewig et al., 2003.澳际中介
References and citation
When citing the system, the vectors pXNgate21-3HA, pX-NubWTgate, pNubWT-Xgate and pMetYCgate, and the control constructs plea refer to the publication Obrdlik et al., 2004, PNAS 101; 12242-12247.
When citing the vector pNXgate33-3HA, plea refer to C. Cappellaro and E. Boles, University of Frankfurt, unpublished.
Differences to the vectors published in Obrdlik et al., 2004
The vectors pXNgate21-3HA and pNXgate33-3HA are bad on the corresponding vectors pXNgate and pNXgate in Obrdlik et al. (2004). Their additional feature is the triple HA tag, which allows the construction of ORF-NubG-3HA and NubG-ORF-3HA fusions. The triple HA tag is readily detectable with anti-HA antibodies. In addition, pNXgate33-3HA is a low-copy CEN plasmid and produces significantly higher signal/noi ratio when compared to pNXgate in Obrdlik et al. (2004).
Plea note that the control construct KAT1-N has not a triple but single HA-tag peptide fud to the C-terminus!
