Table 1. A. thaliana PAL gene under investigation in this project.
|
Chromosome
|
Chromosome Locus
|
Clone Locus
|
WSU I.D.
|
Protein Acc #
|
mRNA
|
|
2
|
At2g37040
|
T1J8.22
|
Atpal1
|
AAP59438
|
AY303128
|
|
3
|
At3g10340
|
F14P13.6
|
Atpal4
|
AAP59440
|
AY303130
|
|
3
|
At3g53260
|
T4D2.190
|
Atpal2
|
AAP59439
|
AY303129
|
|
5
|
At5g04230
|
F21E1.150
|
Atpal3
|
AY528562
|
Table 2. Summary of isolated A. thaliana PAL cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
cDNA Source
|
Expression Vector
|
Transformation (E. coli strains)
|
Purified to Homogeneity
|
PAL activity
|
|
At2g37040
|
Atpal1
|
ABRC
|
pET Blue-1
|
Tuner (DE3pLacI)
|
+
|
+
|
| At3g10340 |
Atpal4
|
Plant
|
pET Blue-1 |
Tuner (DE3pLacI)
|
+
|
+
|
| At3g53260 |
Atpal2
|
Plant
|
pET Blue-1 |
Tuner (DE3pLacI)
|
+
|
+
|
| At5g04230 |
Atpal3
|
Plant*
|
pTrcHis/NT TOPO
|
Tuner (DE3pLacI)
|
+
|
+
|
*Entire plant was used for RNA extraction
Table 3. Summary of isolated A. thaliana PAL promoters and progress toward identifying temporal and spatial patterns of expression.
|
Gene
|
WSU #
|
PCR4 vector
|
PCambia 1305.1
|
Agrobacterium tumefaciens
|
Arabidopsis transformation
|
| At2g37040 |
Atpal1
|
+
|
+
|
+
|
+
|
| At3g10340 |
Atpal4
|
+
|
+
|
+
|
+
|
| At3g10340 |
Atpal4
|
+
|
+
|
+
|
+
|
| At3g53260 |
Atpal2
|
+
|
+
|
+
|
+
|
| At5g04230 |
Atpal3
|
+
|
+
|
+
|
+
|
Table 4. A. thaliana 4CL gene under investigation in this project.
|
Chromosome
|
Chromosome Locus
|
Clone Locus
|
WSU I.D.
|
Protein Acc #
|
mRNA
|
|
1
|
At1g51680
|
F19C24.11
|
At4CL1
|
AAQ86588
|
AY376729
|
|
3
|
At3g21240
|
MXL8.10
|
At4CL2
|
AAQ86587
|
AY376728
|
|
1
|
At1g65060
|
F16G16.6
|
At4CL3
|
AAQ86589
|
AY376730
|
|
5
|
At5g63380
|
K9H21.11
|
At4CL4
|
AAQ86590
|
AY376731
|
|
3
|
At3g21230
|
MXL8.9
|
At4CL5
|
AAQ86591
|
AY376732
|
|
4
|
At4g05160
|
C6L9.15
C17L7.80 |
At4CL6
|
AAM67483
|
AY122950
|
|
4
|
At4g19010
|
F13C5.180
|
At4CL7
|
AAQ86592
|
AY376733
|
|
3
|
At3g48990
|
T2J13.170
|
At4CL8
|
AAA61449
|
AY117254
|
|
1
|
At1g20510
|
F5M15.17
|
At4CL9
|
AAK64105
|
AY040047
|
|
1
|
At1g62940
|
F16P17.9
|
At4CL10
|
AAQ86593
|
AY376734
|
|
5
|
At5g38120
|
F16F17.12 MXA21.2
|
At4CL11
|
AAQ86594
|
AY376735
|
Table 5. Summary of isolated A. thaliana 4CL cDNAs and progress toward characterizing the heterologously expressed proteins.
| Gene | WSU # | cDNA Source |
Cloning in TOPO TA
|
Expression Vector (pCRT7-CT)
|
Transformation (E. coli strains)
|
Purified to Homogeneity
|
4CL Activity
|
| At1g51680 | At4CL1 |
whole plant
|
+
|
+
|
+
|
+
|
+
|
| At3g21240 | At4CL2 |
whole plant
|
+
|
+
|
+
|
+
|
+
|
| At1g65060 | At4CL3 |
whole plant
|
+
|
+
|
+
|
+
|
+
|
|
At5g63380
|
At4CL4 |
whole plant
|
+
|
+
|
+
|
+
|
nd
|
|
At3g21230
|
At4CL5 |
whole plant
|
+
|
+
|
+
|
+
|
+
|
|
At4g05160
|
At4CL6 |
ABRC
(U19441) |
+
|
+
|
+
|
+
|
nd
|
|
At4g19010
|
At4CL7 |
whole plant
|
+
|
+
|
+
|
+
|
nd
|
|
At3g48990
|
At4CL8 |
ABRC
(U11110) |
+
|
+
|
+
|
+
|
nd
|
|
At1g20510
|
At4CL9 |
ABRC
(U09223) |
+
|
+
|
+
|
+
|
nd
|
|
At1g62940
|
At4CL10 |
whole plant
|
+
|
+
|
+
|
+
|
+
|
|
At5g38120
|
At4CL11 |
whole plant
|
+
|
+
|
+
|
+
|
nd
|
Table 6. Summary of isolated A. thaliana 4CL promoters and progress toward identifying temporal and spatial patterns of expression.
|
Gene
|
WSU #
|
PCR4 vector
|
pCambia
1305.1 |
Agrobacterium tumefaciens
|
Arabidopsis transformation
|
|
At1g51680
|
At4CL1
|
+
|
+
|
+
|
+
|
|
At3g21240
|
At4CL2
|
+
|
+
|
+
|
+
|
|
At1g65060
|
At4CL3
|
+
|
+
|
+
|
+
|
|
At5g63380
|
At4CL4
|
+
|
+
|
+
|
+
|
|
At3g21230
|
At4CL5
|
+
|
+
|
+
|
+
|
|
At4g05160
|
At4CL6
|
+
|
+
|
+
|
+
|
|
At4g19010
|
At4CL7
|
+
|
+
|
+
|
+
|
|
At3g48990
|
At4CL8
|
+
|
+
|
+
|
+
|
|
At1g20510
|
At4CL9
|
+
|
+
|
+
|
+
|
|
At1g62940
|
At4CL10
|
+
|
+
|
+
|
+
|
|
At5g38120
|
At4CL11
|
+
|
+
|
+
|
+
|
Table 7. A. thaliana CCOMT gene under investigation in this project.
|
Chromosome
|
Chromosome locus
|
Clone locus
|
WSU #
|
Protein Acc #
|
mRNA
|
|
4
|
At4g34050
|
F28A23.190
|
AtCCOMT1
|
||
|
4
|
At4g26220
|
T25K17.30
|
AtCCOMT2
|
||
|
1
|
At1g67990
|
T23K23.16
|
AtCCOMT3
|
||
|
1
|
At1g67980
|
T23K23.17
|
AtCCOMT4
|
||
|
1
|
At1g24735
|
F5A9.20
|
AtCCOMT5
|
Table 8. Summary of isolated A. thaliana CCOMT cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
cDNA source
|
Cloning vector
|
Transformation
(E.coli strain) |
Expression strain
|
Purified to homogeneity
|
CCOMT activity
|
|
At4g34050
|
AtCCOMT1
|
RNA from 2-week seedlings
|
pET-101
pET-151 |
TOP-10
TOP-10 |
|||
|
At4g26220
|
AtCCOMT2
|
RNA from 2-week seedlings
|
pET-101
pET-151 pET-44 |
TOP-10
TOP-10 Novablue |
BL21-Star
|
||
|
At1g67990
|
AtCCOMT3
|
RNA from 2-week seedlings
|
pET-151
|
TOP-10
|
|||
|
At1g67980
|
AtCCOMT4
|
RNA from 2-week seedlings
|
pET-151
|
TOP-10
|
BL21-Star
|
||
|
At1g24735
|
AtCCOMT5
|
RNA from 2-week seedlings
|
pET-151
|
TOP-10
|
Table 9. A. thaliana COMT gene under investigation in this project.
|
Chromosome
|
Chromosome locus
|
Clone locus
|
WSU #
|
Protein Acc #
|
mRNA
|
|
1
|
At5g54160
|
K18G13.3
|
AtCOMT1
|
||
|
1
|
At1g51990
|
F5F19.5
|
AtCOMT2
|
||
|
5
|
At5g53810
|
MGN6.20
|
AtCOMT3
|
||
|
3
|
At3g53140
|
T4D2.70
|
AtCOMT4
|
||
|
5
|
At5g37170
|
MJG14.10
|
AtCOMT5
|
||
|
1
|
At1g63140
|
F16M19.12
|
AtCOMT6
|
||
|
1
|
At1g77520
|
T5M16.11
|
AtCOMT7
|
||
|
1
|
At1g21130
|
T22I11.4
|
AtCOMT8
|
||
|
1
|
At1g77530
|
T5M16.12
|
AtCOMT9
|
||
|
1
|
At1g33030
|
F9L11.18
|
AtCOMT10
|
||
|
1
|
At1g21100
|
T22I11.7
|
AtCOMT11
|
||
|
1
|
At1g21120
|
T22I11.5
|
AtCOMT12
|
||
|
1
|
At1g21110
|
T22I11.6
|
AtCOMT13
|
||
|
1
|
At1g62900
|
F16P17.4
|
AtCOMT14
|
||
|
1
|
At1g76790
|
F28O16.16
|
AtCOMT15
|
||
|
4
|
At4g35150
|
T12J5.20
|
AtCOMT16
|
||
|
4
|
At4g35160
|
T12J5.30
|
AtCOMT17
|
Table 10. Summary of isolated A. thaliana COMT cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
cDNA Source
|
Expression Vector
|
Transformation (E. coli strains)
|
Purified to Homogeneity
|
COMT Activity
|
|
At5g54160
|
AtCOMT1
|
Plant
|
pEt101
|
+
|
+
|
+
|
|
At1g51990
|
AtCOMT2
|
|||||
|
At5g53810
|
AtCOMT3
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At3g53140
|
AtCOMT4
|
ABRC
|
pEt101
|
+
|
+
|
–
|
|
At5g37170
|
AtCOMT5
|
|||||
|
At1g63140
|
AtCOMT6
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g77520
|
AtCOMT7
|
|||||
|
At1g21130
|
AtCOMT8
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g77530
|
AtCOMT9
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g33030
|
AtCOMT10
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g21100
|
AtCOMT11
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g21120
|
AtCOMT12
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g21110
|
AtCOMT13
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At1g62900
|
AtCOMT14
|
|||||
|
At1g76790
|
AtCOMT15
|
Plant
|
pEt101
|
+
|
+
|
–
|
|
At4g35150
|
AtCOMT16
|
|||||
|
At4g35160
|
AtCOMT17
|
Plant
|
pEt101
|
+
|
+
|
–
|
Table 11. A. thaliana CCR gene under investigation in this project.
|
Chromosome
|
Chromosome Locus
|
Clone Locus
|
WSU I.D.
|
Clone Acc #
|
Protein Acc #
|
mRNA
|
|
1
|
At1g15950
|
T24D18.5
|
Atccr1
|
|||
|
1
|
At1g66800
|
F4N21.7
|
Atccr13
|
|||
|
1
|
At1g68540
|
T26J14.11
|
Atccr4
|
|||
|
1
|
At1g76470
|
F14G6.7
|
Atccr9/10
|
|||
|
1
|
At1g80820
|
F23A5.17
|
Atccr2
|
|||
|
2
|
At2g02400
|
T16F16.19
|
Atccr3
|
|||
|
2
|
At2g23910
|
T29E15.11
|
Atccr6
|
|||
|
2
|
At2g33590
|
F4P9.36
|
Atccr8
|
|||
|
2
|
At2g33600
|
F4P9.37
|
Atccr11
|
|||
|
4
|
At4g30470
|
F17I23.190
|
Atccr7
|
|||
|
5
|
At5g14700
|
T15N1.190
|
Atccr5
|
|||
|
5
|
At5g14700
|
T15N1.190
|
Atccr5a
|
|||
|
5
|
At5g58490
|
MQJ2.6
|
Atccr12
|
Table 12. Summary of isolated A. thaliana CCR cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
cDNA Source
|
Expression Vector
|
Transformation (E. coli strains)
|
Purified to Homogeneity
|
CCR Activity
|
|
At1g15950
|
Atccr1
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
BL21* |
+
|
+
|
|
At1g80820
|
Atccr2
|
ABRC
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
+
|
+
|
|
At2g02400
|
Atccr3
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At1g68540
|
Atccr4
|
Plant
|
pET Blue-1
pET101/D |
Tuner (DE3pLacI)
BL21* |
+
|
–
|
|
At5g14700
|
Atccr5
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At5g14700
|
Atccr5a
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At2g23910
|
Atccr6
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At4g30470
|
Atccr7
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At2g33590
|
Atccr8
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At1g76470
|
Atccr9/10
|
|||||
|
At2g33600
|
Atccr11
|
Plant
|
pET Blue-1
pCRT7/CT TOPO |
Tuner (DE3pLacI)
|
||
|
At5g58490
|
Atccr12
|
|||||
|
At1g66800
|
Atccr13
|
Table 13. Summary of isolated A. thaliana CCR promoters and progress toward identifying temporal and spatial patterns of expression.
|
Gene
|
WSU #
|
PCR4 vector
|
pCambia
1305.1 |
Agrobacterium tumefaciens
|
Arabidopsis transformation
|
|
At1g15950
|
Atccr1
|
+
|
+
|
+
|
+
|
|
At1g80820
|
Atccr2
|
+
|
|||
|
At2g02400
|
Atccr3
|
+
|
+
|
+
|
+
|
|
At1g68540
|
Atccr4
|
||||
|
At5g14700
|
Atccr5
|
+
|
|||
|
At5g14700
|
Atccr5a
|
||||
|
At2g23910
|
Atccr6
|
+
|
|||
|
At4g30470
|
Atccr7
|
||||
|
At2g33590
|
Atccr8
|
+
|
+
|
||
|
At1g76470
|
Atccr9/10
|
+
|
|||
|
At2g33600
|
Atccr11
|
+
|
+
|
||
|
At5g58490
|
Atccr12
|
+
|
+
|
+
|
+
|
|
At1g66800
|
Atccr13
|
Table 14. A. thaliana CAD gene under investigation in this project.
|
Chromosome
|
Chromosome
Locus |
Clone Locus
|
WSU #
|
Protein Acc #
|
mRNA Acc #
|
|
1
|
At1g72680
|
F28P22.13
|
AtCAD1
|
AAP40269
|
AY288079
|
|
2
|
At2g21730
|
F7D8.5
|
AtCAD2
|
AAP59430
|
AY302077
|
|
2
|
At2g21890
|
F7D8.21
|
AtCAD3
|
AAP59431
|
AY302078
|
|
3
|
At3g19450
|
MLD14.30
|
AtCAD4
|
AAP59434
|
AY302081
|
|
4
|
At4g34230
|
F10M10.11/
F28A23.10 |
AtCAD5
|
AAP59435
|
AY302082
|
|
4
|
At4g37970
|
F20D10.90
|
AtCAD6
|
AAP59428
|
AY302075
|
|
4
|
At4g37980
|
F20D10.100
|
AtCAD7
|
AAP59432
|
AY302079
|
|
4
|
At4g37990
|
F20D10.110
|
AtCAD8
|
AAP59433
|
AY302080
|
|
4
|
At4g39330
|
T22F8.230
|
AtCAD9
|
AAP59429
|
AY302076
|
Table 15. Summary of isolated A. thaliana CAD cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
CDNA source
|
Expression
Vector cloning |
Transformation
(E. coli strain) |
Purified to
Homogeneity |
|
At1g72680
|
AtCAD1
|
Leaves, Stems,
and Siliques |
pET101/D-TOPO
|
Tuner(DE3)
|
+
|
|
At2g21730
|
AtCAD2
|
Leaves, Stems,
and Siliques |
pET101/D-TOPO
|
Tuner(DE3)
|
+
|
|
At2g21890
|
AtCAD3
|
Leaves, Stems,
and Siliques |
pET101/D-TOPO
|
Tuner(DE3)
|
+
|
|
At3g19450
|
AtCAD4
|
Leaves, Stems,
and Siliques |
pTrcHis2-TOPO
|
TOP10
|
+
|
|
At4g34230
|
AtCAD5
|
Leaves, Stems,
and Siliques |
pTrcHis2-TOPO
|
TOP10
|
+
|
|
4g37970
|
AtCAD6
|
Leaves, Stems,
and Siliques |
pET101/D-TOPO
|
Tuner(DE3)
|
+
|
|
At4g37980
|
AtCAD7
|
Leaves, Stems,
and Siliques |
pTrcHis2-TOPO
|
TOP10
|
+
|
|
At4g37990
|
AtCAD8
|
Leaves, Stems,
and Siliques |
pTrcHis2-TOPO
|
TOP10
|
+
|
|
At4g39330
|
AtCAD9
|
Leaves, Stems,
and Siliques |
pET101/D-TOPO
|
Tuner(DE3)
|
+
|
Table 16. Kinetic parameters for CAD homologues, AtCAD2 – 5, 7 and 8.
|
Substrate
|
Km
(µM) |
Vmax
(pKat/µg) |
kcat
(sec–1) |
kenz
(M–1 sec–1) |
|
|
AtCAD4
|
p-Coumaryl aldehyde
|
47
|
44.0
|
3.44
|
74,000
|
| Caffeyl aldehyde |
87
|
17.1
|
1.33
|
15,000
|
|
| Coniferyl aldehyde |
65
|
26.8
|
2.10
|
32,000
|
|
| 5-OH Coniferyl aldehyde |
85
|
32.3
|
2.52
|
30,000
|
|
| Sinapyl aldehyde |
274
|
9.2
|
0.72
|
2,600
|
|
| ATCAD5 | p-Coumaryl aldehyde |
13
|
187.3
|
14.52
|
1,091,000
|
| Caffeyl aldehyde |
68
|
94.1
|
7.30
|
107,000
|
|
| Coniferyl aldehyde |
35
|
157.4
|
12.19
|
348,000
|
|
| 5-OH Coniferyl aldehyde |
22
|
106.9
|
8.28
|
370,000
|
|
| Sinapyl aldehyde |
20
|
177.0
|
13.72
|
700,000
|
|
| ATCAD2 | p-Coumaryl aldehyde |
114
|
3.3
|
0.27
|
2,400
|
| Caffeyl aldehyde |
161
|
22.2
|
1.81
|
11,000
|
|
| Coniferyl aldehyde |
452
|
8.0
|
0.65
|
1,400
|
|
| 5-OH Coniferyl aldehyde |
336
|
16.4
|
1.34
|
4,000
|
|
| Sinapyl aldehyde |
2161
|
48.1
|
3.93
|
1,800
|
|
| ATCAD3 | p-Coumaryl aldehyde |
292
|
20.1
|
1.65
|
5,600
|
| Caffeyl aldehyde |
581
|
9.7
|
0.80
|
1,400
|
|
| Coniferyl aldehyde |
362
|
4.8
|
0.39
|
1,100
|
|
| 5-OH Coniferyl aldehyde |
534
|
17.9
|
1.47
|
2,800
|
|
| Sinapyl aldehyde |
629
|
9.1
|
0.74
|
1,200
|
|
| ATCAD7 | p-Coumaryl aldehyde |
320
|
28.6
|
2.18
|
6,800
|
| Caffeyl aldehyde |
3685
|
79.0
|
6.04
|
1,600
|
|
| Coniferyl aldehyde |
675
|
13.0
|
0.99
|
1,500
|
|
| 5-OH Coniferyl aldehyde |
756
|
2.7
|
0.21
|
280
|
|
| Sinapyl aldehyde |
313
|
0.6
|
0.05
|
150
|
|
| ATCAD8 | p-Coumaryl aldehyde |
302
|
20.4
|
1.59
|
5,300
|
| Caffeyl aldehyde |
683
|
7.0
|
0.54
|
800
|
|
| Coniferyl aldehyde |
141
|
5.6
|
0.44
|
3,100
|
|
| 5-OH Coniferyl aldehyde |
457
|
10.4
|
0.81
|
1,800
|
|
| Sinapyl aldehyde |
898
|
28.9
|
2.25
|
2,500
|
Table 17. Summary of isolated A. thaliana CAD promoters and progress toward identifying temporal and spatial patterns of expression.
|
Gene
|
WSU #
|
PCR4 vector
|
pCambia
1305.1 |
Agrobacterium tumefaciens
|
Arabidopsis transformation
|
Analysis of T2 plants
|
| At1g72680 | AtCAD1 |
+
|
+
|
+
|
+
|
+
|
| At2g21730 | AtCAD2 |
+
|
+
|
+
|
+
|
–
|
| At2g21890 | AtCAD3 |
+
|
+
|
+
|
+
|
+
|
| At3g19450 | AtCAD4 |
+
|
+
|
+
|
+
|
+
|
| At4g34230 | AtCAD5 |
+
|
+
|
+
|
+
|
+
|
| At4g34230 | AtCAD5 |
+
|
+
|
+
|
+
|
+
|
| At4g34230 | AtCAD5 |
+
|
+
|
+
|
+
|
+
|
| At4g34230 | AtCAD5 |
+
|
+
|
+
|
+
|
+
|
| At4g37970 | AtCAD6 |
+
|
+
|
+
|
+
|
–
|
| At4g37980 | AtCAD7 |
+
|
+
|
+
|
+
|
–
|
| At4g37990 | AtCAD8 |
+
|
+
|
+
|
+
|
+
|
| At4g39330 | AtCAD9 |
+
|
+
|
+
|
+
|
+
|
Table 18. A. thaliana APDBR gene under investigation in this project.
|
Chromosome
|
Chromosome
Locus |
Clone Locus
|
WSU #
|
Protein Acc #
|
mRNA Acc #
|
|
1
|
At1g65560
|
F5I14.9
|
AtAPDBR9
|
||
|
3
|
At3g03080
|
T17B22.23
|
AtAPDBR5
|
||
|
3
|
At3g59845
|
F24G16.1
|
AtAPDBR8
|
||
|
5
|
At5g16970
|
F2K13.120
|
AtAPDBR1
|
||
|
5
|
At5g16990
|
F2K13.140
|
AtAPDBR2
|
||
|
5
|
At5g17000
|
F2K13.150
|
AtAPDBR3
|
||
|
5
|
At5g16980
|
F2K13.130
|
AtAPDBR4
|
||
|
5
|
At5g38000
|
K18L3.2
/ K19A23.1 |
AtAPDBR6
|
||
|
5
|
At5g37940
|
K18L3.100
|
AtAPDBR6-1
|
||
|
5
|
At5g16960
|
F2K13.110
|
AtAPDBR7
|
||
|
5
|
At5g37980
|
K18L3.140
|
AtAPDBR10
|
Table 19. Summary of isolated A. thaliana APDBR cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
cDNA source
|
Expression
Vector |
Transformation
(E. coli strain) |
Purified to
Homogeneity |
APDBR
Activity |
|
At1g16970
|
AtAPDBR 1
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
+
|
|
At2g16990
|
AtAPDBR 2
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
–
|
|
At2g17000
|
AtAPDBR 3
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
–
|
|
At3g16980
|
AtAPDBR 4
|
—
|
Underway
|
|||
|
At4g03080
|
AtAPDBR 5
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
–
|
|
At4g38000
|
AtAPDBR 6
|
Leaves, Stems,
and Siliques |
+
|
Tuner(DE3)
|
Underway
|
Underway
|
|
At4g37940
|
AtAPDBR 6-1
|
Leaves, Stems,
and Siliques |
+
|
Tuner(DE3)
|
Underway
|
Underway
|
|
At4g16960
|
AtAPDBR 7
|
Leaves, Stems,
and Siliques |
+
|
Tuner(DE3)
|
Underway
|
Underway
|
|
At4g59845
|
AtAPDBR 8
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
+
|
|
At4g65560
|
AtAPDBR 9
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
+
|
|
At4g37980
|
AtAPDBR 10
|
Leaves, Stems,
and Siliques |
+
|
TOP10
|
+
|
–
|
Table 20. Summary of isolated A. thaliana APBDR promoters and progress toward identifying temporal and spatial patterns of expression.
|
Gene
|
WSU #
|
PCR4 vector
|
pCambia
1305.1 |
Agrobacterium tumefaciens
|
Arabidopsis transformation
|
|
At1g16970
|
AtAPDBR 1
|
+
|
+
|
+
|
+
|
|
At2g16990
|
AtAPDBR 2
|
+
|
+
|
+
|
+
|
|
At2g17000
|
AtAPDBR 3
|
+
|
+
|
+
|
+
|
|
At3g16980
|
AtAPDBR 4
|
+
|
+
|
–
|
–
|
|
At4g03080
|
AtAPDBR 5
|
+
|
+
|
+
|
+
|
|
At4g38000
|
AtAPDBR 6
|
+
|
+
|
–
|
–
|
|
At4g37940
|
AtAPDBR 6-1
|
+
|
+
|
–
|
–
|
|
At4g16960
|
AtAPDBR 7
|
+
|
+
|
+
|
+
|
|
At4g59845
|
AtAPDBR 8
|
+
|
+
|
+
|
+
|
|
At4g65560
|
AtAPDBR 9
|
+
|
+
|
+
|
+
|
|
At4g37980
|
AtAPDBR 10
|
+
|
+
|
+
|
+
|
Table 21. A. thaliana DIR gene under investigation in this project.
|
Chromosome
|
Chromosome
Locus |
Clone Locus
|
WSU #
|
Protein Acc #
|
mRNA Acc #
|
|
5
|
At5g42510
|
MDH9.21
|
ATDIR1
|
||
|
5
|
At5g42500
|
MDH9.20
|
ATDIR2
|
||
|
5
|
At5g49040
|
K19E20.19
|
ATDIR3
|
||
|
2
|
At2g21110
|
F26H11.13
|
ATDIR4
|
||
|
1
|
At1g64160
|
F22C12.8
|
ATDIR5
|
||
|
4
|
At4g23690
|
F9D16.160
|
ATDIR6
|
||
|
3
|
At3g13650
|
MMM17.15
|
ATDIR7
|
||
|
3
|
At3g13662
|
MMM17.6
|
ATDIR8
|
||
|
2
|
At2g39430
|
F12L6.9
|
ATDIR9
|
||
|
2
|
At2g28670
|
T8O18.4
|
ATDIR10
|
||
|
1
|
At1g22900
|
F19G10.14
|
ATDIR11
|
||
|
4
|
At4g11180
|
T22B4.160
|
ATDIR12
|
||
|
4
|
At4g11190
|
T22B4.170
|
ATDIR13
|
||
|
4
|
At4g11210
|
T22B4.190 F8L21.1
|
ATDIR14/16
|
||
|
4
|
At4g38700
|
T9A14.4
F20M13.260 |
ATDIR15
|
||
|
4
|
At4g13580
|
T6G15.130
|
ATDIR18
|
Table 22. Summary of sense and antisense AtDIR plants.
|
Gene
|
WSU #
|
Sense (S)/
Antisense (AS) Construct |
To seeds
|
T1 seeds
|
T2 seeds
|
T3 plants for analysis
|
|
At5g42510
|
ATDIR1
|
S
|
+
|
|||
|
AS
|
+
|
+
|
||||
|
At5g42500
|
ATDIR2
|
S
|
+
|
+
|
+
|
|
|
AS
|
+
|
+
|
+
|
|||
|
At5g49040
|
ATDIR3
|
S
|
+
|
|||
|
AS
|
+
|
+
|
||||
|
At2g21110
|
ATDIR4
|
S
|
+
|
+
|
+
|
|
|
AS
|
+
|
+
|
+
|
|||
|
At1g64160
|
ATDIR5
|
S
|
+
|
+
|
+
|
+
|
|
AS
|
+
|
+
|
+
|
+
|
||
|
At4g23690
|
ATDIR6
|
S
|
+
|
+
|
||
|
AS
|
+
|
+
|
+
|
|||
|
At3g13650
|
ATDIR7
|
S
|
+
|
+
|
||
|
AS
|
+
|
|||||
|
At3g13662
|
ATDIR8
|
S
|
+
|
+
|
+
|
|
|
AS
|
+
|
|||||
|
At2g39430
|
ATDIR9
|
S
|
+
|
+
|
+
|
+
|
|
AS
|
+
|
+
|
+
|
+
|
||
|
At2g28670
|
ATDIR10
|
S
|
+
|
+
|
||
|
AS
|
+
|
+
|
+
|
+
|
||
|
At1g22900
|
ATDIR11
|
S
|
+
|
|||
|
AS
|
+
|
+
|
||||
|
At4g11180
|
ATDIR12
|
S
|
+
|
+
|
||
|
AS
|
+
|
+
|
||||
|
At4g11190
|
ATDIR13
|
S
|
+
|
+
|
+
|
|
|
AS
|
+
|
+
|
+
|
|||
|
At4g11210
|
ATDIR14/16
|
S
|
+
|
+
|
+
|
+
|
|
AS
|
+
|
+
|
+
|
+
|
||
|
At4g38700
|
ATDIR15
|
S
|
+
|
|||
|
AS
|
+
|
+
|
||||
|
At4g13580
|
ATDIR18
|
S
|
+
|
+
|
+
|
+
|
|
AS
|
+
|
+
|
Table 23. Summary of isolated A. thaliana PER cDNAs and progress toward characterizing the heterologously expressed proteins.
|
Gene
|
WSU #
|
cDNA source
|
Expression vector
|
Transformation (Drosophila)
|
Activity
|
|
At5g17820
|
AtPER1
|
30 d root
|
pIB
|
||
|
At3g03670
|
AtPER2
|
14 d whole plants
|
pIB
|
||
|
At4g26010
|
AtPER3
|
ABRC
|
pMT
|
||
|
At5g06720
|
AtPER4
|
||||
|
At4g11290
|
AtPER5
|
ABRC
|
|||
|
At5g22410
|
AtPER6
|
ABRC
|
|||
|
At5g64100
|
AtPER7
|
ABRC
|
pMT
|
||
|
At5g39590
|
AtPER8
|
pMT
|
|||
|
At1g05260
|
AtPER9
|
30 d root
|
pIB
|
||
|
At5g06730
|
AtPER10
|
30 d root
|
pIB/ pMT
|
||
|
At1g34330
|
AtPER11
|
||||
|
At3g32980
|
AtPER12
|
ABRC
|
|||
|
At2g18980
|
AtPER13a
|
ABRC
|
|||
|
At1g71695
|
AtPER13b
|
ABRC
|
|||
|
At3g49960
|
AtPER14
|
||||
|
At1g49660
|
AtPER15a
|
pIB/ pMT
|
|||
|
At3g48690
|
AtPER15b
|
||||
|
At3g21770
|
AtPER16
|
ABRC
|
|||
|
At5g19890
|
AtPER17
|
ABRC
|
|||
|
At5g40150
|
AtPER19
|
||||
|
At2g35380
|
AtPER20
|
30 d root
|
pIB
|
||
|
At4g17690
|
AtPER21
|
||||
|
At4g16270
|
AtPER22
|
||||
|
At4g33870
|
AtPER23
|
||||
|
At3g17070
|
AtPER24
|
||||
|
At2g34060
|
AtPER25
|
||||
|
At2g37130
|
AtPER26
|
ABRC
|
|||
|
At1g05250
|
AtPER27
|
ABRC
|
|||
|
At5g64120
|
AtPER28
|
ABRC
|
|||
|
At5g67400
|
AtPER29
|
||||
|
At5g14130
|
AtPER30
|
ABRC
|
|||
|
At1g14480
|
AtPER31
|
||||
|
At4g31760
|
AtPER32
|
||||
|
At3g50990
|
AtPER33
|
||||
|
At2g41480
|
AtPER34
|
30 d root
|
pMT
|
||
|
At5g24070
|
AtPER35
|
30 d root
|
pMT/pIB
|
||
|
At5g58400
|
AtPER36
|
||||
|
At2g38390
|
AtPER37
|
||||
|
At2g38380
|
AtPER38
|
ABRC
|
|||
|
At4g37520
|
AtPER39
|
30 d root
|
pIB |
||
|
At4g30170
|
AtPER40
|
30 d root
|
|||
|
At5g66390
|
AtPER41
|
||||
|
At1g68850
|
AtPER42
|
||||
|
At4g08770
|
AtPER43
|
ABRC
|
|||
|
At1g44970
|
AtPER44
|
||||
|
At2g22420
|
AtPER45
|
||||
|
At4g33420
|
AtPER46
|
30 d root
|
pIB
|
||
|
At5g64110
|
AtPER47
|
||||
|
At5g58390
|
AtPER48
|
30 d root
|
pIB
|
||
|
At5g05340
|
AtPER49
|
||||
|
At1g24110
|
AtPER50
|
||||
|
At1g14490
|
AtPER51
|
30 d root
|
pIB
|
||
|
At2g43480
|
AtPER52
|
ABRC
|
|||
|
At4g21960
|
AtPER53
|
ABRC
|
|||
|
At3g49120
|
AtPER54
|
ABRC
|
|||
|
At3g49110
|
AtPER55
|
||||
|
At5g42180
|
AtPER56
|
ABRC
|
|||
|
At3g01190
|
AtPER57
|
30 d root
|
pIB
|
||
|
At5g51890
|
AtPER58
|
ABRC
|
|||
|
At1g30870
|
AtPER59
|
||||
|
At3g28200
|
AtPER60
|
ABRC
|
|||
|
At2g18140
|
AtPER61
|
||||
|
At4g36430
|
AtPER62
|
ABRC
|
|||
|
At5g47000
|
AtPER63
|
||||
|
At5g15180
|
AtPER64
|
ABRC
|
|||
|
At2g39040
|
AtPER65
|
||||
|
At1g77100
|
AtPER66
|
||||
|
At2g24800
|
AtPER67
|
||||
|
At1g34510
|
AtPER68
|
||||
|
At5g19880
|
AtPER69
|
||||
|
At5g06720
|
AtPER70
|
ABRC
|