Gene List on a Plant Tumor-inducing Plasmid, pTi-SAKURA in ...
Transcript of Gene List on a Plant Tumor-inducing Plasmid, pTi-SAKURA in ...
DNA RESEARCH 7, 371-380 (2000) Record
Gene List on a Plant Tumor-inducing Plasmid, pTi-SAKURA inAgrobacterium tumefadens MAFF301001
Yoshiyuki HATTORI,1'* Misugi URAJI,1^ Katsunori SUZUKI,1'* Nobuyuki OHTA,1 Kumi IWATA,1
Akira KATO,2 and Kazuo YOSHIDA1'*
Department of Biological Science, Graduate School of Science, Hiroshima University, Higashi-Hiroshima739-8526, Japan1 and Hokkaido Agricultural Experiment Station, Sapporo 061-8555, Japan2
(Received 2 October 2000; revised 11 October 2000)
Plant infectious Agrobacteria harbor large plasmids Tior Ri (about 200 kb), which induce crown gall or hairyroot syndromes,1 respectively. These plasmids have threeimportant functional regions, transfer DNA (T-DNA)region, virulence (Vir) region and conjugation transfer(Tra) region. The T-DNA region is integrated into plantchromosomes, and the subsequent expression of its plantgrowth hormone genes causes the formation of a crowngall or hairy root tumor.2 The Vir region is specificallyresponsible for processing T-DNA and transfer into plantchromosomes.3 The Tra region is involved in bacterialconjugation. These gene systems have provided the fun-damental bases of plant genetic engineering. Recently,we have succeeded in the first complete sequencing of aTi plasmid and its short report has been published.4 Inthis paper, we describe the list of genes on the plasmid,pTi-SAKURA in A. tumefadens with a detailed map,available database information and codon usage.
Table 1 shows a list of 195 genes including hypotheticalgenes and other significant sequences on pTi-SAKURAwith their function, GC% and similarity against proteinsequences in the public databases. Here, the hypothet-ical genes indicate ORFs which were not identified indatabases or had very low similarity. Although the cir-cular map and the functional classification of these ORFswere described in the previous paper,4 the detailed loca-tion map is shown in Fig. 1 in which each color indicatesthe similar function of genes.
Codon usages of total 195 genes including hypothet-ical genes are summarized in Table 2. As shown inTable 2, arginine codons (7.9%) are relatively rich af-ter the commonly abundant alanine (10.9%) and leucine(9.9%) codons. A preliminary analysis of 19 proba-ble chromosomal genes of Agrobacterium in the DNADatabase showed lower arginine codons (6.3%), and al-
Communicated by Masahiro Sugiura* To whom correspondence should be addressed. Department
of Biological Science, Graduate School of Science, HiroshimaUniversity, Kagamiyama 1-3-1, Higashi-Hiroshima 739-8526,Japan, Tel. +81-0824-24-7455, Fax. +81-0824-24-0734, E-mail:kyoshida@sci. hiroshima-u. ac. j p
t Equally contributed authors
most the same alanine (11.2%) and leucine (9.1%) codonusage. The frequent use of arginine codons in pTi-SAKURA probably indicate the abundant supply of argi-nine from plant crown galls in the form of nopaline. Ter-mination codon usage in pTi-SAKURA is summarized asfollows: the plasmid's genes used 52% UGA, 25% UAGand 23% UAA as termination codons while the chromo-somal genes used 74% UGA, 16% UAG and 11% UAA.
Acknowledgements: We are grateful to the editorand anonymous reviewers for their suggestion.
References
1. Zambryski, P., Tempe, J., and Schell, J. 1989, Transferand function of T-DNA genes from Agrobacterium Ti andRi plasmids in plants, Cell, 56, 193-201.
2. Jayaswal, R. K., Veluthambi, K., Gelvin, S. B., andSlightom, J. L. 1987, Double-stranded cleavage of T-DNAand generation of single-stranded T-DNA molecules inEscherichia colt by a wrD-encoded border-specific en-donuclease from Agrobacterium tumefadens, J. Bacteriol,169, 5035-5045.
3. Rogowsky, P. M., Powell, B. S., Shirasu, K., Lin, T.-S., Morel, P., Zyprian, E. M., Steck, T. R., and Kado,C. I. 1990, Molecular characterization of the vir regulonof Agrobacterium tumefadens: Complete nucleotide se-quence and gene organization of the 28.63-kbp reguloncloned as a single unit, Plasmid, 23, 85-106.
4. Suzuki, K., Hattori, Y., Uraji, M., Ohta, N., Iwata, K.,Murata, K., Kato, A., and Yoshida, K. 2000, Completenucleotide sequence of a plant tumor-inducing Ti plas-mid, Gene, 242, 331-336.
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
Tab
le 1
. G
ene
list
of p
Ti-
SAK
UR
A.
to
tiorf
* F
eatu
re
Nuc
leot
ides
No
No.
st
art
(end
)-en
d (s
tart
)S
tran
d G
ene
nam
eFu
nctio
n**
GC
con
tent
(%
) Si
mila
rity
(%
) ag
ains
t pr
otei
ns
Tra
nscr
ipti
onal
reg
ulat
ion
Nop
alin
e tr
ansp
ort
Nop
alin
e tr
ansp
ort
Nop
alin
e tr
ansp
ort
: no
palin
e-bi
ndin
g pr
otei
nN
opal
ine
tran
spor
tT
rans
crip
tion
al r
egul
ator
for
nop
alin
e ca
tabo
lism
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
Con
juga
tion
: au
toin
duce
r sy
nthe
sis
Reg
ulat
ory
sequ
ence
for
con
juga
tion
Reg
ulat
ory
sequ
ence
for
con
juga
tion
Veg
etat
ive
DN
A r
eplic
atio
nV
eget
ativ
e D
NA
rep
licat
ion
Ori
gin
of v
eget
ativ
e D
NA
rep
licat
ion
Veg
etat
ive
DN
A r
eplic
atio
n(D
NA
inv
erta
se)
Plas
mid
sta
bili
tyT
rans
posi
tion
(D
NA
inv
erta
se)
tiorf
l***
tiorf
2tio
rf3
tiorf
4tio
rf5
tiorf
6tio
rf7
tiorf
8tio
rf9
tiorf
lOti
orfl
1ti
orfl
2ti
orfl
3ti
orfl
4ti
orfl
5ti
orfl
6ti
orfl
7ti
orfl
8
tior
fl9
tiorf
20
tiorf
21tio
rf22
tiorf
23tio
rf24
tiorf
25tio
rf26
tiorf
27tio
rf28
tiorf
29tio
rf30
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
UT
RU
TR
CD
SC
DS
rep-
orig
inC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
S
1559
- 24
7925
94-
3319
3321
- 40
3141
06-
4957
5022
- 57
9560
16-
6951
7308
- 86
2486
37-
9113
9113
- 99
6799
85-1
0647
1066
2-11
825
1185
2-12
070
1206
7-12
870
1284
2-15
310
1532
1-15
620
1561
3-16
014
1600
4-16
975
1697
2-17
607
1766
8-17
681
1775
9-17
772
1797
1-19
188
1941
8-20
428
2048
3-20
641
2064
3-21
962
2197
1-22
912
2319
4-24
306
2443
1-24
985
2509
9-25
434
2603
1-26
600
2668
2-27
899
2800
7-28
723
2873
2-29
316
2948
4-30
311
HG
—
nocM
—
nocQ
- no
cVno
cP+
raoc
Rtr
bltr
bHtr
bGtr
bFtr
bhtr
bKtr
bitr
bEtr
bF)
trbC
trbB
tral
tra
box
tra
box
+ re
pA+
repB
oriV
+
repC
—
ivrl
+
HG
+ H
G+
stb-
h+
iv
rZ+
H
G+
HG
+ H
G4-
H
G
57.0
57.6
56.7
55.6
57.8
54.7
60.4
65.6
62.6
62.0
64.5
69.4
60.2
62.9
65.0
62.9
63.4
60.0
53.9
57.0
49.7
58.6
61.9
61.5
60.4
56.9
59.7
59.4
59.8
60.3
60.1
29.9
(B
. su
btili
s al
s R
pro
tein
)10
0 (p
TiC
58 N
ocM
)10
0 ( P
TiC
58 N
ocQ
)99
.3 (
pTiC
58
Noc
T)99
.2 (
pTiC
58 N
ocP
)10
0 ( P
TiC
58 N
ocK
)79
.3 (
PT
il59
55 7
V6I
)71
.5 (
PT
il59
55 7
VfcH
)84
.5 (
PT
il59
55 7
V6G
)93
.2 (
PT
il59
55 T
rbF
)82
.7 (
pTil
5955
7V
6L)
46.7
(pT
il59
55 7
V6K
)83
.3 (
pTil
5955
7V
6J)
88.4
(PT
il59
55 7
V6E
)81
.8 (
PT
il59
55 7
V6D
)72
.6 (
PT
il59
55 7
V6C
)88
.2 (
PT
il59
55 7
V6B
)99
.5 (
pTiC
58 T
ral)
96.0
(PT
iB6S
3 re
pA)
80.1
(PT
iB6S
3 re
pB)
75.3
(pT
iB6S
3 re
pC)
65.0
(pN
GR
234a
y4c
G)
42.7
(pN
GR
234a
y4c
F)
33.7
(T.
Jer
roox
idan
s)44
.0 (
P.
aeru
gino
sa)
72.3
(pN
GR
234a
j/^
RN
)
o B CD O H
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o OS
Tab
le 1
. C
ontin
ued.
tiorf
31tio
rf32
tiorf
33tio
rf34
tiorf
35tio
rf36
tiorf
37tio
rf38
tiorf
39tio
rf40
tiorf
41tio
rf42
tiorf
43tio
rf44
tiorf
45tio
rf46
tiorf
47tio
rf48
tiorf
49tio
rf50
tiorf
51tio
rf52
tiorf
53tio
rf54
tiorf
55tio
rf56
tiorf
57tio
rf58
tiorf
59tio
rf60
tiorf
61tio
rf62
tiorf
63tio
rf64
tiorf
65
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
S
3057
1-30
813
3084
2-31
234
3123
8-32
401
3246
8-33
610
3376
7-34
333
3518
9-36
184
3629
2-36
918
3773
2-38
499
3871
5-39
128
3934
7-40
270
4023
1-40
827
4026
7-40
872
4082
4-41
285
4150
8-42
695
4165
8-42
260
4227
3-42
674
4280
4-47
885
4504
8-45
719
4807
0-49
983
4833
1-49
263
5000
8-50
466
5050
6-51
522
5166
0-52
295
5234
1-53
321
5335
9-54
531
5452
4-55
558
5560
5-56
216
5621
3-56
785
5680
3-57
648
5854
3-59
142
5890
7-59
914
5944
5-60
371
6037
1-61
309
6167
5-62
364
6239
5-63
135
+ H
G+
HG
- H
G-
HG
- H
G+
in
t-
muf
T-h
+ H
G-
HG
+ H
G-
HG
+ H
G-
HG
- H
G+
HG
+ H
G+
HG
- H
G+
HG
- H
G+
HG
+ H
G—
no
dQ-h
- H
G-
HG
- H
G-
HG
- ri
bk-h
- H
G+
HG
+ H
G+
HG
+ H
G-
HG
- H
G
(DN
A i
nteg
rase
)
DN
A m
ethy
latio
n
Inte
ract
ion
with
pla
nt
(GT
P cy
cloh
ydro
lase
)
51.0
58.0
56.2
58.0
60 0
57.4
54.2
51.3
60.4
64.6
62.8
62.5
63.4
62.5
62.7
60.0
63.7
61.8
63.1
62.1
64.3
54.5
50.5
49.1
49.5
51.9
51.1
48.2
52.6
66.2
63.4
61.3
61.0
63.8
59.2
55.6
(pN
GR
234a
y^R
C)
43.0
{B
. su
btil
is M
utV
hom
olog
ous
yjhB
)
50.0
(C
. bu
met
ii 1
52aa
hyp
othe
tica
l pr
otei
n)
42.8
(C
. bu
met
ii Q
pHl
hypo
thet
ical
pro
tein
)
29.2
(B
. st
earo
ther
mop
hilu
s B
ssH
l)
58.0
(R
hizo
bium
BR
816
Nod
Q p
rote
in)
33.0
(M
. tu
becu
losi
s R
ibA
)
42.0
(pN
GR
234a
y4e
C)
50
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Tab
le 1
. C
ontin
ued.
tiorf
66tio
rf67
tiorf
68tio
rf69
tiorf
70tio
rf71
tiorf
72tio
rf73
tiorf
74tio
rf75
tiorf
76tio
rf77
tiorf
78tio
rf79
tiorf
80ti
orfg
ltio
rf82
tiorf
83tio
rf84
tiorf
85tio
rf86
tiorf
87tio
rf88
tiorf
89tio
rf90
tiorf
91tio
rf92
tiorf
93tio
rf94
tiorf
95tio
rf96
tiorf
97tio
rf98
tiorf
99tio
rflO
O
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
S
6323
4-63
698
6381
0-64
088
6410
6-64
600
6459
7-65
064
6527
3-66
733
6678
1-67
158
6792
6-68
207
6828
0-69
194
6941
0-69
886
6994
7-70
543
7146
9-71
784
7175
5-72
201
7234
1-72
685
7270
6-73
749
7288
3-73
482
7417
2-74
477
7497
3-75
548
7554
5-77
695
7852
7-79
258
7925
1-79
919
7997
9-80
443
8069
5-81
771
8187
3-82
364
8260
6-83
526
8363
2-84
138
8425
0-85
098
8518
8-86
156
8624
9-87
133
8741
3-88
168
8824
2-89
252
8935
6-90
369
9047
5-91
557
9155
4-92
339
9245
6-93
784
9381
2-94
450
- H
G-
HG
- H
G-
HG
- H
G+
H
G+
H
G-
HG
- H
G-
HG
+
HG
- H
G+
H
G+
H
G-
HG
- H
G+
H
G+
H
G+
li
gE+
H
G+
H
G+
H
G-
HG
+
HG
+
HG
- H
G+
qo
r+
H
G+
H
G-
HG
+
HG
+
HG
+
HG
+
HG
+
HG
Lig
nin
degr
adat
ion
(qui
none
oxi
dore
duct
ase)
53.8
46.6
56.6
53.2
58.2
55.0
50.7
57.6
54.5
63.0
63.8
61.7
57.1
58.3
58.8
56.2
59.7
59.6
56.8
54.4
57.9
54.5
56.3
57.3
56.4
60.4
58.9
58.2
60.6
51.3
54.7
54.3
57.8
54.6
52.9
50.0
(pN
GR
234a
y4e
B)
38.0
(Sp
hing
omon
as l
igE
bet
a-et
hera
se)
o (I a 53'
H >
41.0
(E
. co
li q
or p
rote
in)
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
o
Tab
le 1
. C
ontin
ued.
tiorf
lOl
tior
fl02
tior
fl03
tior
fl04
tior
fl05
tior
fl06
tior
fl07
tior
fl08
tior
fl09
tior
fllO
tior
f lll
tior
fll2
tior
fll3
tior
fll4
tior
fll5
tior
fll6
tior
fll7
tior
fllS
tior
fll9
tior
fl20
tior
fl21
tior
fl22
tior
fl23
tior
fl24
tior
fl25
tior
fl26
tior
fl27
tior
fl28
tior
fl29
tior
fl30
tior
fl31
tior
fl32
tior
fl33
tior
fl34
tior
fl35
tior
fl36
tior
fl37
tior
fl38
tior
fl39
tior
fl40
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
U'l'
Kre
peat
-uni
tC
DS
CD
SC
DS
CD
SU
TR
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
UT
RC
DS
CD
SC
DS
CD
SC
DS
UT
RC
DS
CD
SC
DS
CD
SC
DS
CD
SU
TR
CD
S
9458
4-95
198
9550
0-95
778
9591
8-96
175
9660
6-96
932
9731
0-97
711
9773
2-99
681
9966
8-99
883
9988
8-10
0184
1UU
261-
1U02
7410
0345
-100
360
1004
41-1
0402
510
4022
-104
552
1044
76-1
0572
310
5737
-106
363
1065
84-1
0659
710
6888
-107
196
1073
55-1
0907
910
9182
-109
511
1095
41-1
1024
511
0463
-111
971
1119
72-1
1355
511
4166
-114
942
1150
00-1
1656
511
6599
-117
564
1175
61-1
1858
011
8577
-119
503
1194
96-1
2035
612
0372
-121
172
1212
16-1
2198
312
2359
-123
444
1231
99-1
2410
712
4442
-124
454
1245
84-1
2575
912
5919
-127
094
1273
70-1
2829
312
8377
-128
646
1291
89-1
3012
713
0303
-130
316
1305
78-1
3100
613
1028
-131
462
1314
62-1
3182
113
1906
-132
181
1341
07-1
3481
713
5962
-136
693
1374
12-1
3742
613
7530
-140
031
- H
G—
hu
pT-
HG
+ H
G-
HG
- fr
aG-
traD
- tr
aCtr
a B
oxor
iT i
nver
ted
+ tr
aA+
tr
ap+
traB
+ H
G tra
Box
+ tr
aM—
m
cpA
- H
G-
fraR
- H
G-
HG
-r-
accR
+ ac
cA+
accB
4-
accC
+ ac
cD+
accE
+ ac
cF(p
de)
+ ac
cG(a
pp)
+ H
G-
tps
vir
Box
+ pi
nFl
4-
pmF2
+ tp
s-h
- H
G+
inrF
vir
Box
+ H
G+
virK
+
HG
- H
G-
HG
- tz
svi
r B
ox+
virA
Hyd
roge
n-up
take
Con
juga
tion
Con
juga
tion
Con
juga
tion
Reg
ulat
ory
sequ
ence
for
con
juga
tion
repe
at
Con
juga
tion
(ori
gin
of t
rans
fer)
Con
juga
tion
Con
juga
tion
Con
juga
tion
Reg
ulat
ory
sequ
ence
for
con
juga
tion
Con
juga
tion
Che
mor
ecep
tor
Tra
nscr
ipti
onal
act
ivat
or f
or c
onju
gatio
nSu
cros
e ph
osph
olyl
atio
n (p
hosp
holy
lase
)
Rep
ress
or f
or a
ce g
ene
expr
essi
onA
groc
inop
ine
tran
spor
tA
groc
inop
ine
tran
spor
tA
groc
inop
ine
tran
spor
tA
groc
inop
ine
tran
spor
tA
groc
inop
ine
tran
spor
tA
groc
inop
ine
cata
boli
sm (
phos
phod
iest
eras
e)A
groc
inop
ine
cata
boli
sm (
phos
phat
e ph
osph
atas
e)
Tra
nspo
sitio
n (t
rans
posa
se)
Reg
ulat
ory
sequ
ence
for
vir
ulen
ceV
irul
ence
Vir
ulen
ceT
rans
posi
tion
(tra
nspo
sase
)
Vir
ulen
ceR
egul
ator
y se
quen
ce f
or v
irul
ence
.Vir
ulen
ce
Cyt
okin
in s
ynth
esis
(pr
enyl
tra
nsfe
rase
)R
egul
ator
y se
quen
ce f
or v
irul
ence
Vir
ulen
ce
62.0
59.9
60.5
56.9
59.2
65.2
64.4
66.3
63.4
65.0
65.5
61.6
54.7
59.9
52.1
58.7
55.2
58.9
57.0
54.8
58.5
58.0
56.8
56.9
56.8
55.7
53.9
57.8
53.5
58.1
54.4
61.5
51.0
55.2
51.0
58.1
65.2
56.4
54.0
52.3
98.0
(pT
iC58
hup
T pr
otei
n)
82.0
(pT
iRlO
tra
G p
rote
in)
78.0
(pT
iC58
tra
D p
rote
in)
76.0
(pT
iRlO
tra
C p
rote
in)
72.0
(pT
iC58
tra
A p
rote
in)
72.0
(pT
iC58
tra
F pr
otei
n)93
.0 (
pTiC
58 t
raB
pro
tein
)
99.0
(pT
iC58
tra
M p
rote
in)
62.0
(pT
il59
55 o
rfi
prot
ein)
99.0
(pT
iC58
tra
R p
rote
in)
74.0
(A
. vi
tis)
99.0
(pT
iC58
acc
R)
99.0
(pT
iC58
acc
A)
42.0
(B
. su
btili
s op
pD)
45.0
(B
. su
btili
s dp
pF)
36.0
(B
. su
btili
s dp
pD)
36.0
(E
. co
li dp
pC)
30.0
(E
. co
li su
hB)
32 (
E.
coli
ugpQ
)
99 (
pTiC
58)
78 (
PT
iA6
pmF
l)84
(PT
iA6
pmF2
)
100
(Pro
babl
e vi
rF)
100
(Pro
babl
e vi
rK)
100
(pT
iC58
tzs
)
99.7
(PT
iC58
vir
A)
X
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
Tab
le 1
. C
ontin
ued.
tior
fl41
tior
fl42
tior
fl43
tior
fl44
tior
fl45
tior
fl46
tior
fl47
tior
fl48
tior
fl49
tior
fl50
tior
fl51
tior
fl52
tior
fl53
tior
fl54
tior
fl55
tior
fl56
tior
fl57
tior
fl58
tior
fl59
tior
fl60
tior
fl61
tior
fl62
tior
fl63
tior
fl64
tior
fl65
tior
fl66
tior
fl67
tior
fl68
tior
fl69
tior
fl70
tior
fl71
tior
fl72
tior
fl73
tior
fl74
tior
fl75
tior
fl76
tior
fl77
tior
fl78
tior
fl79
tior
fl80
UT
RC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
UT
RC
DS
CD
SC
DS
UT
RC
DS
CD
SC
DS
CD
SU
TR
CD
SC
DS
UT
RC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
rep
orig
inC
DS
CD
SC
DS
repe
at-u
nit
CD
SC
DS
repe
at-u
nit
CD
SC
DS
CD
SC
DS
1404
42-1
4045
514
0576
-141
313
1413
13-1
4167
814
1678
-142
004
1420
04-1
4437
314
4390
-145
052
1451
53-1
4604
014
6076
-146
243
1462
30-1
4694
314
6940
-147
821
1478
18-1
4895
114
8992
-150
026
1500
68-1
5008
115
0245
-151
006
1511
48-1
5175
615
1759
-152
454
1525
95-1
5258
215
2725
-153
168
1532
02-1
5454
515
4560
-156
581
1566
55-1
5866
115
8554
-158
567
1587
73-1
6129
816
1675
-162
001
1622
59-1
6227
216
2387
-162
638
1627
54-1
6294
516
2959
-164
629
1647
44-1
6675
016
7260
-167
889
1679
18-1
6864
916
8646
-169
200
1691
90-1
6936
316
9404
-170
180
1701
73-1
7054
117
0755
-170
988
1709
35 1
7095
917
1321
-172
436
1733
40-1
7402
317
4479
-175
663
1759
38-1
7596
417
6176
-177
225
1780
97-1
7854
017
8602
-178
629
1798
21-1
8051
918
1225
-181
917
1823
96-1
8334
018
3802
-184
908
vir
Box
virB
lvi
rB2
virB
3vi
rBA
mrB
5vi
rBG
virB
lvi
rB8
virB
9vi
rBW
inrB
llvi
r B
oxvi
rGvi
rC2
tnrC
lvi
r B
oxvi
rDl
virD
2vi
rD3
virD
Avi
r B
oxO
RF
5vi
rHvi
r B
oxH
Gvi
rEl
virE
2H
GH
GH
GH
GH
Gac
cGac
cF'
HG
LB acs
5-h
HG
IR IS o
rfIS
orf
IR rolB
-h21
-he ac
s
Reg
ulat
ory
sequ
ence
for
vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Reg
ulat
ory
sequ
ence
for
vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Reg
ulat
ory
sequ
ence
for
vir
ulen
ceV
irul
ence
Vir
ulen
ceV
irul
ence
Vir
ulen
ceR
egul
ator
y se
quen
ce f
or v
irul
ence
Vir
ulen
ceV
irul
ence
Reg
ulat
ory
sequ
ence
for
vir
ulen
ce
Vir
ulen
ceV
irul
ence
(ind
ucib
le b
y ac
etos
yrin
gone
)
accG
gen
e ho
mol
og(a
ccF
gene
fra
gmen
t)
Ter
min
atio
n of
T D
NA
tra
nsfe
rT
-DN
A g
ene
: ag
roci
nopi
ne s
ynth
esis
T-D
NA
gen
e
Inse
rtio
n se
quen
ce t
erm
inal
rep
eat
Inse
rtio
n se
quen
ce t
erm
inal
rep
eat
T-D
NA
gen
e (r
ootin
g lo
ci)
T-D
NA
gen
eT
-DN
A g
ene
T-D
NA
gen
e (a
groc
inop
ine
synt
hesi
s)
56.4
58.0
56.9
55.3
56.7
54.2
55.4
55.0
54.1
57.9
55.4
53.7
55.8
54.0
54.1
58.3
55.8
53.1
55.2
54.7
55.9
45.8
50.3
51.9
58.9
60.0
58.7
59.8
58.7
57.5
53.4
49.6
48.1
50.1
63.0
62.6
50.1
50.7
50.5
46.1
99.5
(PT
iC58
99.7
(PT
iC58
100
(pT
iC58
99.6
(PT
iC58
99.8
(pT
iC58
99.7
(pT
iC58
100
(pT
iC58
99.3
(pT
iC58
99.9
(pT
iC58
100
(pT
iC58
99.3
(PT
iC58
virB
l)m
rB2)
mrB
3)m
rB4)
virB
b)vi
rB6)
virB
7)vi
rB8)
virB
9)vi
rBlO
)m
r Bll
)
100
( PT
iC58
vir
G)
100
( PT
iC58
wrC
2)10
0 ( P
TiC
58
virC
l)
100
(pT
iC58
uir
Dl)
99.7
(PT
iC58
vir
D2)
99.2
(PT
iC58
«rD
3)99
.4 (
PT
iC58
t;ir
D4)
99.2
(PT
iC58
orf
i)98
.2 (
pTiC
58 v
irH
)
100
(pT
iC58
vi
rEl)
99.7
(PT
iC58
t;ir
E2)
75.4
(pT
iC58
occ
G)
72.7
(pT
iC58
acc
F)
71.4
(pR
iA4
TL
-DN
A
OR
Fl)
48.6
(pT
iTm
4 5
gene
pro
duct
)
89.1
(PT
iTm
4 /S
866
OR
Fl)
64.1
(PT
iTm
4 75
866
OR
F4)
26.9
(pR
iA4
rolB
)34
.1 (
pTil
5955
hyp
othe
tica
l pr
otei
n 21
)98
.7 (
pTiC
58 h
ypot
heti
cal
prot
ein)
99.4
(pT
iC58
agr
ocin
opin
e sy
ntha
se)
o n H In
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
Z o
Tab
le 1
. C
ontin
ued.
tior
fl81
tior
fl82
tior
fl83
tior
fl84
tior
fl85
tior
fl86
tior
fl87
tior
fl88
tior
fl89
tior
fl90
tior
fl91
tior
fl92
tior
fl93
tior
fl94
tior
fl95
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
Sre
p-or
igin
CD
SC
DS
CD
SC
DS
CD
SC
DS
CD
SC
DS
1856
72-1
8633
718
6844
-188
247
1885
87-1
9085
419
1551
-192
273
1927
89-1
9334
919
3832
-194
455
1954
45-1
9668
619
7026
-197
050
1971
96-1
9825
819
8306
-199
388
1994
65-2
0043
920
0487
-201
905
2019
02-2
0219
520
2192
-203
310
2039
19-2
0595
520
5959
- 14
35**
+
5-
tms2
+
tmsl
+
tmr
+
6a-
6b—
no
sR
B—
oc
d-
odh
—
arc
—
noxA
- H
G—
no
xB+
hy
uA-h
! +
hy
uB-h
T-D
NA
gen
e 45
.1T
-DN
A g
ene
(tum
or m
orph
olog
y, i
ndol
acet
amid
e hy
drol
ase)
51
.2T
-DN
A g
ene
(tum
or m
orph
olog
y, t
rypt
opha
n 2-
mon
ooxy
gena
se)
50.0
T-D
NA
gen
e (t
umor
mor
phol
ogy,
iso
pent
enyl
tra
nsfe
rase
) 50
.4T
-DN
A g
ene
48.8
T-D
NA
gen
e 48
.7T
-DN
A g
ene
(nop
alin
e sy
nthe
sis)
48
.6O
rigi
n of
T-D
NA
tra
nsfe
rN
opal
ine
cata
bolis
m (
orni
thin
e cy
clod
eam
inas
e)
58.7
Nop
alin
e ca
tabo
lism
(op
ine
dehy
drog
enas
e)
59.7
Nop
alin
e ca
tabo
lism
(ar
gina
se)
60.7
Nop
alin
e ca
tabo
lism
(no
palin
e de
hydr
ogen
ase)
63
.359
.5N
opal
ine
cata
bolis
m (
nopa
line
dehy
drog
enas
e)
61.0
(hyd
anto
inas
e)
61.4
(hyd
anto
inas
e)
60.3
81.5
(pT
iCG
474
5 ge
ne p
rodu
ct)
93.8
(pT
iTm
4 in
dola
ceta
mid
e hy
doro
lase
)92
.3 (
pTiA
6NC
try
ptop
han
2-m
onoo
xyge
nase
)10
0 (p
TiT
37 i
sope
nten
yl t
rans
fera
se)
98.9
(pT
iT37
pro
tein
6a)
100
(pT
iT37
hyp
othe
tica
l pr
otei
n 6b
)97
.5 (
pTiT
37 n
opal
ine
synt
hase
)
99.4
(pT
iC58
orn
ithi
ne c
yclo
deam
inas
e)91
.2 (
pTiC
58 h
ypot
heti
cal
40kd
pro
tein
)10
0 (p
TiC
58 a
rgin
ase)
99.3
(pT
iC58
Nox
A)
98.9
(pT
iC58
Nox
B)
34.0
(P
seud
omon
as h
ydan
toin
uti
liza
tion
pro
tein
A)
31.0
(P
seud
omon
as h
ydan
toin
uti
liza
tion
pro
tein
B)
X CD
*tio
rf i
ndic
ates
an
OR
F of
pT
i-SA
KU
RA
. C
odin
g se
quen
ces
wer
e ta
ken
to b
e O
RFs
if
they
wer
e pr
edic
ted
by G
eneM
ark
anal
ysis
and
sim
ilar
to t
hose
in
data
base
s in
bot
hle
ngth
and
hom
olog
y. E
ven
non-
hit
sequ
ence
s w
ere
take
n t o
be
OR
Fs if
the
y w
ere
long
er t
han
80 c
odon
s an
d ha
d pr
omot
er a
nd r
ibos
ome-
bind
ing-
like
sequ
ence
s, (
see
ref.
4 in
deta
il)**
The
fun
ctio
nal
prod
uct
with
add
itio
nal
info
rmat
ion
is s
how
n in
par
enth
esis
.**
*tio
rfl9
5 is
loc
ated
ove
r th
e st
arti
ng c
oord
inat
e A
, w
hich
is
the
5'-p
rotr
udin
g en
d ba
se b
y E
coR
l cl
eava
ge s
ite (
see
Fig.
1).
Abb
revi
atio
ns i
ndic
ate:
CD
S; p
redi
cted
cod
ing
sequ
ence
, re
p-or
igin
; re
plic
atio
n or
igin
, re
peat
-uni
t; r
epea
t un
it,
UT
R;
untr
ansc
ribe
d re
gion
, H
G;
hypo
thet
ical
gen
e, R
B a
ndL
B;
righ
t bo
rder
, le
ft b
orde
r, B
. su
btil
is;
Bac
illu
s su
btil
is,
T.
ferr
ooxi
dans
; T
hiob
acil
lus
ferr
ooxi
dans
, P
. ae
rugi
nosa
; P
seud
mon
as a
erug
inos
a, C
. bu
rnet
ii;
Cox
iell
a bu
rnet
ii,
B.
stea
roth
erm
ophi
lus;
Bac
illu
s st
earo
ther
mop
hilu
s, M
. tu
berc
ulos
is;
Myc
obac
teri
um t
uber
culo
sis,
E.
coli
; E
sche
rich
ia c
oli,
A.
viti
s; A
grob
acte
rium
vit
is.
The
seq
uenc
e da
ta o
f pT
i-SA
KU
RA
has
bee
n de
posi
ted
in t
he D
DB
J/G
enB
ank/
EM
BL
dat
abas
e un
der
the
acce
ssio
n N
o. A
B01
6260
.
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
CO 00
(hyu
B)
tiorf
l95
(noc
K)
tiorf
6
trab
oxor
iV
(rep
A)
(rep
B)
tiorf
l 9
tiorf
20(r
epC
)tio
rQl
tiorf
23 t
iorf
24
lbtio
rfl
tiorC
tio
rB
tiorf4
tio
rfS
(noc
M)(
nocQ
)(no
cT)
(nod
?)tio
rf7
tiorf
? tio
rfl
1 tio
rt13
tiorf
l4
tiorf
l5 t
iorf
l7(t
rbV
) (t
rbG
) (t
rbV
) (t
rbf)
(t
rbE
) (t
rbD
) (t
rbB
)tio
rfS
tiorf
l 0 t
iorf
l 2
tiorf
l 6
tiorf
l 8(t
rbH
) (t
rbF
) (t
rbK
) (t
rbC
) {t
ral)
tior£
22{i
wl)
25kb
(ivr
2)(s
tb)
tiorf
26
tiorf
28tio
rf25
tio
rf27
tio
rf29
tio
rBO
tiorB
l tiorf
32(i
nl)
tiorf
36tio
rf38
tiorf
42
tiorf
46tio
rf40
tio
rf45
(met
hyla
se)
tiorf
47tio
rf49
O CD
tiorf
33
tiorB
4tio
rG5
25kb
tiorf
37(m
utT
)tio
rf39
tiorf
41
tiorf
44tio
rf43
tiorf
48tio
rf50
50kb
tiorf
52tio
rf51
tior
fld
tiorf
Bl
tiorff
iO
>
tior
%3
tiorf
72
tiorf
74tio
rf71
tio
rf73
tior
f76
tiorf
78 t
iorf
79
t
H in >
50kb
tiorf
53
tiorf
54 t
iorfS
5(n
odQ
)tio
rf56
tior
f57
tiorf
59tio
rf58
(rib
A)
tiorf
i54
tiorf
66 t
iorf
68
tiorf
70tio
rf65
tio
rf67
tio
rfB
9tio
rf75
tio
rf77
tiorf
SO
ti
orf
Sl
75kb
tiorf
82
tiorf
83
iorf8
5
tiorf
86tio
rf87
tiorf
90tio
rf89
(qor
)tio
rf92
tio
rf93
tio
rf94
tiorf
96
tiorf
9710
>tio
rf99
tio
rfl 0
0tio
rfl 0
4
tiorf
88tio
rf91
tiorf
95
75kb
tiorf
l 01
tiorf
l 03
tiorf
l 02
(hup
T)
tiorf
l 05
tiorf
l 06
tiorf
l 08
(tra
G)
(tra
C)
tiorf
l 07
(tra
D)
100k
b
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
oriT
trab
oxib
oi(t
raA
)ti
orfl
09
(trd
B)
iorf
lll
tiorf
l 10
^io
rfl
trab
ox (tra
M)
tiorf
l 13
2
(acc
A)
tiorf
l 20
(acc
R)
tiorf
l 19
(acc
B)
tiorf
l 21
(acc
C)
tiorf
l 22
(acc
E)
tiorf
l 24
(acc
D)
(acc
F)tio
rfl 2
3 tio
rfl 2
5(acc
G)
tiorf
l 26
vir
box
tiorf
l 27
o
100k
btio
rfl
14
tiorf
l 16
tiorf
l 17
tio
rfl 1
8(m
cpA
) (i
raR
) (p
hosp
holy
lase
)tio
rfl
15
tiorf
l 28
(tps
)12
5kb
vir
box
(pin
Vl)
(tp
s)tio
rfl
30
tiorf
l 31
(vir?)vi
r bo
x ( vi
rK)
lii.ir
l'135
vir
box
tior
fnui
ofl
34 ti
orfl
36
(vir
A)
tiorf
l 40
vir
box
(wV
B2)
tiorf
l 42
(w>
BI)
(v;>
B3)
(v
/rB
4)tio
rfl 4
1 tio
rfl 4
1 tio
rfl
44
(v/>
B6)
(w
>B8)
tiorf
l 46
tiorf
l 48
(v/z
-BIO
)ti
or f
l 50
(w"r
B5)
tiorf
l 4S
(v;V
B7)
(w
>B9)
tinr
fl47
tin
rfl4
Q(v
irB
11)
tinr
flS
l
125k
btio
rfl 3
2ti
orfl
37tio
rfl 3
8tio
rfl
39(t
zs)
150k
b
vir
box
(vir
G)
liorf
I52
vir
box
vir
box
r/VD
I)
(wV
D2)
iorf
l 55
tio
rfl
56(v
/rD
3)tio
rfl
57(v
/>D
4)tio
rfl
58(O
RF
5)
Iior
fl59
vir
box (w
rEl)
liorf
l62(
w>E
2)to
rfl6
1 tio
rll 6
3tio
rfl
64LB
(5 g
ene)
tiorf
l 73
iiorf"
153
(v/>
C2)
150k
b tio
rfl 5
4(v
i/C
l)
tiorf
l 60
tior
fl65
tio
rfl 6
7 ti
ortl
69ti
orfl
71 t
iorf
l 72
tiorf
l 66
(acc
G)
(acs
)tio
rfl 6
8 tio
rfl 7
0(a
ct?)
tiorf
l 74
175k
b
IS(r
oIB
) (2
1 ge
ne)
(e g
ene)
tiorf
l 77
tio
rfl
78
tiorf
l 79
(S g
ene)
tiorf
l 81
(tm
s\)
tiorf
l 83
(tm
r) (
6a g
ene)
tiorf
l 84
tio
rfl 8
5(n
os)
tiorf
l 87
R
B
175k
b
tiorf
l 75
tiorf
l 76
(OR
Fl)
(O
RF
4)tio
rfl 8
0(a
cs)
tiorf
l 82
(tm
sl)
tiorf
l 86
(6b
gene
)
200k
b
tiorf
l 91
tiorf
l 93
(nux
A)
(nox
B)
tior
fl 9
2
(hyu
A)
(hyu
B)
tior
fl94
tio
rfl 9
5
2064
79/1
b
tiorf
l 88
tio
rfl8
9(o
cd)
(arc
)
200k
b
Fig
ure
1.
Det
aile
d ge
ne m
ap o
f pT
i-SA
KU
RA
. G
ene
orga
niza
tion
is e
xpre
ssed
in
a lin
ear
form
. H
oriz
onta
l lin
es s
how
the
ent
ire
leng
th 2
06,4
79 b
p of
pT
i-SA
KU
RA
fro
mri
ght
to le
ft an
d fro
m t
op
to d
own
dire
ctio
n. T
he
star
ting
bas
e is
th
e 5'
-pro
trud
ing
end
base
£co
RI
clea
vage
site
on
tior
fl.9
5. E
ach
vert
ical
lin
e w
hich
cro
sses
th
e ho
rizo
ntal
line
indi
cate
s 5
kb in
dis
tanc
e. E
ach
arro
w i
ndic
ates
the
dir
ectio
n an
d si
ze o
f ea
ch g
ene.
Eac
h O
RF
is n
umbe
red
on o
r un
der
the
arro
w.
Iden
tifie
d ge
nes
are
pare
nthe
size
d.T
he f
unct
iona
lly
rela
ted
gene
s ar
e sh
own
in t
he s
ame
colo
r as
fol
low
s:
gree
n fo
r T
-DN
A;
pink
for
vir
ulen
ce i
nclu
ding
int
erac
tion
with
pla
nts;
red
for
mod
ific
atio
n an
dre
com
bina
tion;
and
blu
e fo
r ve
geta
tive
DN
A r
eplic
atio
n.
The
pos
ition
s of
fun
ctio
nal
sequ
ence
s, s
uch
as o
riT
, or
iV,
tra
box,
vir
box
are
indi
cate
d by
ver
tical
arr
ows,
(se
eT
able
1)
Downloaded from https://academic.oup.com/dnaresearch/article-abstract/7/6/371/471117by gueston 19 March 2018
380 Gene List of pTi-SAKURA
Table 2. Codon usage in 195 ORFs of pTi-SAKURA.
First Second
[Vol. 7,
UUU(F) 795
UUC(F) 1414
UUA(L) 219
UUG(L) 819
UCU(S) 400
UCC(S) 745
UCA(S) 428
UCG(S) 994
UAU(Y) 698
UAC(Y) 567
UAA(-) 44
UAG(-) 49
UGU(C) 182
UGC(C) 408
UGA(-) 102
UGG(W) 739
CUU(L) 1108
CUC(L) 1530
CUA(L) 300
CUG(L) 1689
CCU(P) 493
CCC(P) 598
CCA(P) 537
CCG(P) 1232
CAU(H) 677
CAC(H) 582
CAA(Q) 725
CAG(Q) 1283
CGU(R) 700
CGC(R) 1544
CGA(R) 582
CGG(R) 984
AUU(I) 836
AUC(I) 2000
AUA(I) 371
AUG(M) 1299
ACU(T) 370
ACC(T) 1040
ACA(T) 516
ACG(T) 1077
AAU(N) 708
AAC(N) 920
AAA(K) 759
AAG(K) 1266
AGU(S) 347
AGC(S) 896
AGA(R) 330
AGG(R) 414
GUU(V) 902
GUC(V) 1705
GUA(V) 392
GUG(V) 1037
GCU(A) 941
GCC(A) 2119
GCA(A) 1120
GCG(A) 2060
GAU(D) 1570
GAC(D) 1644
GAA(E) 1673
GAG(E) 1773
GGU(G) 868
GGC(G) 1945
GGA(G) 798
GGG(G) 721
Amino acids are expressed by one-letter code in parenthesis.(-) indicates stop codons.
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