... by introducing
an NsiI site at the translation start point of galA and by then
cloning a fragment, starting at this NsiI site and containing
the galA gene and approximately 700 bp 3¢-flanking ... galacto-oligosaccharides.
ACKNOWLEDGEMENTS
The authors thank Simon Flitter for the identification and isolation
of the galA containing phage clones and Matthew Illsley for the
analysis of a-
L
-1,5-arabinofuranosidase activity inthe ... demonstrates theimportant role of GALA in
the degradation of both linear and
L
-arabinose-substituted
galactan side chains of pectin. This is in agreement with a
previous study in which the synergy...
... indicated by an arrow inthe right lane. (B) In vitro
bovine trypsin inhibition by the recombinant NtKTI1 protein (open
circles). Soybean trypsin inhibitor (triangles) was assayed in parallel
as ... 4B).
Tobacco plants overexpressing NtKTI1 show
enhanced resistance to R. solani infection
To evaluate thein planta role of NtKTI1 in defense,
sense and antisense lines under the control of the cauli-
flower ... trypsin inhibitor with antifungal activity
from Nicotiana tabacum, plays animportant role in
tobacco’s defense response
Hao Huang*, Sheng-Dong Qi*, Fang Qi, Chang-Ai Wu, Guo-Dong Yang and Cheng-Chao...
... its
NH
2
-terminal region, as well as an Mre11-binding
domain and an ATM-binding domain in its COOH-
terminal region (Fig. 1). FHA and BRCT domains are
often present in eukaryotic nuclear proteins involved
in ... MRN foci in cells exposed to IR, led to the
hypothesis that the MRN complex functions in the
sensing of DNA strand breakage and inthe activation
of the ATM-dependent DNAdamage signaling path-
way ... substrates inthe ATM-depend-
ent DNAdamage signaling pathway [10,11,39].
Given that ATM is a central playerinthe cellular
response to DNA strand breakage, it is important to
understand the mechanisms...
... of
the N-terminal region on the molecular surface. Inthe wild-
type enzyme, the side chain of Asp266 interacted with the
amino group of the main chain at Glu268. The oxygen of
the main chain ... conformational changes
on the surface. On the other hand, anionic surfactants such
as SDS and LAS bind to the surface regions of protein, and
the surface regions would be unfolded and removed from
the core ... from the
amino acid sequence alignment of ALP I and the neutral
subtilisins. These candidate residues did not occur at the
corresponding positions of the alkaline subtilisins. Fur-
thermore, on the...
... spans the entire thickness
of the inner retina. The Mu
¨
ller cell constitutes an ana-
tomical link between the retinal neurones and supports
their activities by exchanging molecules between the
other ... RPE65c and GS staining (B3), and 4¢,6-diamidino-2-phenylindole (DAPI) staining (B4), respectively. (B5-8) High magnification
images of the boxed area in (B4) showing RPE65c staining (B5), GS staining ... contribute to
providing chromophore to melanopsin to maintain
their light sensitivities. The expression and function of
RPE65c in other cell types inthe retina remains to be
investigated.
Earlier...
... linked hairpins inthe membrane
[10]. Importantly, the ion-binding site is not conserved
in hairpin one. If one assumes the same number of
transmembrane helices in V
0
and F
0
, the rotor of
eukaryal ... (lanes 6, 8 and 9), and
individual subunits were detected by silver
staining (lane 6) or immunoblotting using an
antibody against subunit c
1
(lanes 7 and 8)
or subunit c
2 ⁄ 3
(lane 9). The antibody
against ... handedness, and two
of the rings face inthe opposite direction in the
membrane to the other two, forming the same pattern
as inthe AFM surface representation of Fig. 2A. By
comparison with the...
... rectangles, b-strands as arrows. ÔdÕ indicates residues inthe dimer interface and ÔtÕ residues in tetramer interface.
Residues lining the active site cavity are underlined. Residues binding ... of
the corresponding intermediate in transketolase [37] and the
model derived for ScPDC [38] (Fig. 6). Inthe immediate
vicinity of the modelled a-carbanion/enamine, there are a
number of invariant ... by amino acids with a large
hydrophobic side chain. Furthermore, there are significant
structural differences between ZmPDC and IPDC on
the one hand, and ScPDC on the other hand involving
the C-terminal...
... patterning in early
embryonic development and inthe antimicrobial immune responsein larvae
and adults. We have investigated the function of Spa
¨
tzle in a lepidopteran
insect, Manduca sexta, in ... the protein was transferred to a
poly(vinylidene difluoride) membrane and stained with 40%
methanol containing 0.025% Coomassie Brilliant Blue
R-250. After destaining with 50% methanol, the band
corresponding ... gene. Inthe clade including
Spa
¨
tzle-1, the branch lengths are noticeably longer and
the bootstrap values are lower than inthe other clades
containing Spa
¨
tzle-2 to Spa
¨
tzle-6, indicating...
... protein incubated with increas-
ing concentrations of either ATP or ATPcS was centri-
fuged followed by assaying the protein concentrations
in the supernatant as well as the pellet. Inthe set ... con-
taining ATPcS, the entire protein sample was recov-
ered inthe supernatant (Fig. 5A) and no protein was
detected inthe pellet fractions (data not shown). In
the same conditions the ATP ... have investigated the changes associ-
ated with the molecular aggregates of mismatch repair
proteins MutS, MutL in relation to their interaction
with mismatch containing DNA and the ongoing ATP
hydrolysis....
... cell
speci®c anticancer therapy [29], we encountered large
differences inthe responses of the various human cancer
cell lines to the same ODN. We have examined this effect in
detail and extended the analysis ... is inactive inthein vitro RNase
H assay, or that some other enzymatic activity is measured.
The detection of increased activity inthe transfectants
overexpressing the coding region of the ... as the ODN h ybridizes to the
center of the input target RNA. The asterisk indicates the input target
RNA; the arrow indicates the digested product bands. The amount of
digested product obtained...
... pUZ8002 bearing the cosmid
with oriT and the S. coelicolor DdesA mutant, single cross-
over exconjugants were screened for kanamycin resistance.
Kanamycin-resistant colonies were isolated and analysed ... As the
DdesA mutant is apramycin resistant, the kanamycin
resistance marker was cloned into an XbaI site of the new
construct (pSKdesA). A 5723-bp XhoI+NotI fragment
containing the desA gene and ... procedures), and the DdesA mutation was veri-
fied in one of the mutants by PCR and Southern blot
analysis. A 1462-bp PCR band corresponding to the
extended resistance cassette was found only in the
mutant...
... narrower line-widths than any
other residues inthe protein. This indicates that the
PEST sequence is an unstructured, highly mobile
region within SOCS3.
The PEST sequence is an insertion inthe SH2
domain
Analysis ... C-terminal
suppressor of cytokine signalling (SOCS) box. SOCS3 and the related pro-
tein, cytokine-inducible SH2-containing protein, are unique among the
SOCS family of proteins in containing a region of mostly ... the kinase inhibitory region and the entire extended SH2
subdomain form a single a-helix. The mapping of the true SH2 domain,
and the location of its C terminus more than 50 residues further...
... mRNA insta-
bility and the involvement of the ARE in this process
[10,11]. In addition, another destabilizing sequence was
identified within the coding region of IFN-b mRNA [12,13].
Whereas the independent ... 1A). We then analysed, by Northern blot, the induc-
tion and decay of IFN-b transcript inthe same conditions.
AsshowninFig.1B,IFN-b mRNA was detectable 4 h
after the beginning of the infection, ... encoding the HA epitope was
inserted at the end of the IFN-b coding sequence to
distinguish the products resulting from the expression of the
DNA constructs and the endogenous gene (Fig. 2). These
constructs...
... been
proposed as a Mg
2+
-binding ligand in a product com-
plex, in addition to its initial role in direct ligation of
the sugar OH group inan analogous manner to that
found intheDNA ⁄ RNA polymerase ... identified Lys19 inthe Haemo-
philus ducreyi CNS, and Lys21 and Arg12 in CNS
from Escherichia coli (Fig. 1) as important catalytic
residues, indicating a role in binding the nucleotide
into the active ... of the purified
F192A and F193A mutant enzymes were obtained in
the same way as the wild-type enzyme (Table 1). The
results support the role of these residues in binding
Neu5Ac, because the mutants...
... dolphins and whales are the closest
divergences before the ruminants, we further examined
the size of the a-chain in whales and dolphins as well
as other ruminants (cattle and goat) to determine the
possible ... time of the tandem repeat evolution in deer
Hp. Interestingly, the inserted panel of Fig. 10 shows
that the a-chains of all the ruminants tested are the a2
type, except for dolphins (n = 5) and whales ... linkages within the a-chain (Cys34 and
68 and Cys93 and 127), and the one at the C-terminal
region (Cys131) cross-links with the b-chain (Cys105) to
form a basic a-b unit. Under such an arrangement,
Cys15...