... Petersburg,
FL, USA) followed by PCR with Taq DNA polymerase
(Promega, Madison, WT, USA) using the primers
5¢-CACACTACACTGGGAAGCAGAG ACTCCAGC-3¢
and 5¢-AGCTGGCCCAGCAGCCATACACAGTTAAAG-
3¢. The cDNA was subcloned ... [43–49].
Acknowledgements
The authors thank Drs Masato Yasui, Sadakazu Aiso
and Masaaki Matsuoka for support; Dr Andrew
P. McMahon for providing the full-length mou-
se Shh...
... earlier maximal pathway
activation and an increase in signal amplitude [3,4].
Furthermore, alterations in the activity of a kinase or
phosphatase may affect the speed of signalling pathway
activation. ... Tamada Y, Nakao K, Nagayama Y, Nakata K,
Ichikawa T, Kawamata Y, Ishikawa H, Hamasaki K,
Eguchi K & Ishii N (2002) p48 Overexpression
enhances interferon-mediated expression and...
... genome with a
GTG start. Sense primer, 5¢-GAGAGACATATGCCA
TCGGCCATTTTAAAG-3¢; antisense primer, 5¢ -GAGA
GAAAGCTTCTAACGCAGGGAGAAAAC-3¢. An alter-
native start site would be the in-frame ATG, 30 nucleotides
downstream ... incor-
porate suitable restriction sites (NdeI-HindIII) into the
5¢- and 3¢ ends of the product. Sense primer, 5¢-GAGA
GAGGATCCATATGACAAAAACCGAGTTAC-3¢, NdeI
site; anti...
... L, Hamid Q & Elias JA (2004) Acidic
mammalian chitinase in asthmatic Th2 inflammation
and IL-13 pathway activation. Science 304, 1678–
1682.
4 Kasprzewska A (2003) Plant chitinases ) regulation ... chitinases
have catalytic domains that are at least as large as
those of the plant enzymes and that may contain at
least six subsites [26,30]. However, the catalytic
domains of Chi...
... wavelength of 280 nm and an
emission wavelength of 400 nm. Adenine at concentrations
ranging from 1 to 1200 pmol was used as a standard for
calculation of the amount of adenine in the samples.
Assay for ... Isolation and enzymatic characterization of lamjapin, the first
ribosome-inactivating protein from cryptogamic algal plant
(
Laminaria japonica
A)
Ren-shui Liu
1
,...
... EC
U
ịC
L
EC
U
ịC
U
2ị
Enzyme activity assays
$
D
-Ala-
D
-Ala carboxypeptidase (CPase) activity was deter-
mined by uorescence-based detection of
D
-Ala in micro-
titer plate-based assays as described in detail previously
[15,22]. ... of
90 min.
D
-Ala product was detected with QuantaBlu
TM
assays. j,
Boc-Cbz-KAA; r, Boc-Ac-KAA; s, Ac-C-KAA; n, Ac
2
-KAA; h,
Boc-H-KAA; d, Ac
2
-KA-
D
-...
... 5¢-G
GTGTTTTATATGGATTAAAAACAATATCACTTAC-3¢.
The mutant plasmid was confirmed by DNA sequencing
before transformation into E. coli strain BL21(DE3). The
expression and purification of the Y211F mutant were ... such
as aromatic amino acids, folic acid and ubiquinone
[3]. The shikimate pathway is crucial in algae, higher
plants, bacteria, apicomplexan parasites and fungi, but
is absent i...