參考文獻
S. enterica serovar Typhimurium LT2
1 kb
tus fumC fumA
K. pneumoniae MGH78578
rstA rstB
ydgC ydgB ydgI
ydgH rstA HP rstB tus fumC fumA
ydgC ydgB ydgI
ydgH HP tus fumC fumA
E. coli K-12 MG1655
rstA rstB ydgB
ydgI
ydgH tus
ydgC
fumC fumA manA ydgA
rstA ompN
S. enterica serovar Typhimurium LT2
1 kb
tus fumC fumA
圖一. 不同菌種中 rstA/rstB 及其上下游基因分析。比較克雷白氏肺炎桿菌、
（A）
ydgB ydgC rstA HP rstB tus
2171 bp
717 bp Deletion
RstA04 RstAp01
ydgB ydgC rstA HP rstB tus
2171 bp
wild type form
mutant form
wild type form
mutant form
1.0
（B）
ydgB ydgC rstA HP rstB tus
3154 bp
1299 bp Deletion
RstA19 RstAp18
ydgB ydgC rstA HP rstB tus
3154 bp
wild type form
mutant form
wild type form
mutant form
圖二. 確認rstA及rstB基因缺損株。（A）利用引子RstA04、RstAp01經PCR增 幅後，以DNA電泳確認。野生株可增幅2171 bp，而rstA缺損株則可增幅1454 bp。Lane 1：野生株，Lanes 2：rstA缺損株，Lanes 3：帶有rstA缺損的質體。
（B）利用引子YWp18，YWp19經PCR後，再以DNA電泳確認。野生株增幅 3154 bp，而rstB缺損株則可增幅1855 bp， Lane 1：野生株，Lanes 2~3：rstB 缺損株，Lanes 4：帶有rstB缺損的質體。
（A） rstA mutant
LB M9 rstA mutant
LB M9
（A） Sulfamethoxazole (25 μg) Sulfonamides Polymyxin B (300 units)
Polymyxin B
Cell Membrane Disruption
20.0 ± 0 20.0 ± 0
19.6 ± 0.2 16.0 ± 0
17.2 ± 0.5 Nalidixic acid (30 μg)
34.6 ± 0.1
Quinolones and Fluoroquinolones
Nucleic Acid Synthesis inhibition
8.0 ± 0 8.6 ± 0.5
8.6 ± 0.5 22.6 ± 0.5
23.0 ± 0.7 Tetracyclin (30 μg)
Tetracyclines Kanamycin (30 μg)
Aminoglycosides Protein Synthesis inhibition
32.0 ± 0
Cefotaxime (30 μg) Cephalosporins Methicillin (5 mcg)
20.6 ± 0.5 Ampicillin (10 μg)
Penicillins)
Cell Wall Synthesis Inhibition
pHY112
Klebsiella pneumoniae CG43S3 Drug Sulfamethoxazole (25 μg) Sulfonamides Polymyxin B (300 units)
Polymyxin B
Cell Membrane Disruption
20.0 ± 0 20.0 ± 0
19.6 ± 0.2 16.0 ± 0
17.2 ± 0.5 Nalidixic acid (30 μg)
34.6 ± 0.1
Quinolones and Fluoroquinolones
Nucleic Acid Synthesis inhibition
8.0 ± 0 8.6 ± 0.5
8.6 ± 0.5 22.6 ± 0.5
23.0 ± 0.7 Tetracyclin (30 μg)
Tetracyclines Kanamycin (30 μg)
Aminoglycosides Protein Synthesis inhibition
32.0 ± 0
Cefotaxime (30 μg) Cephalosporins Methicillin (5 mcg)
20.6 ± 0.5 Ampicillin (10 μg)
Penicillins)
Cell Wall Synthesis Inhibition
pHY112
Klebsiella pneumoniae CG43S3 Drug
（B）
23.2 ± 0.3 22.8 ± 0.2
Cefmetazole (30 μg)
6.0 ± 0 6.0 ± 0
Methicillin (5 mcg)
18.0 ± 0.2 17.0 ± 0.1
Carbenicillin (100 μg)
6.0 ± 0 6.0 ± 0
Sulfamethoxazole (25 μg) Sulfonamides
Antimetabolites
13.8 ± 0.4 13.6 ± 0.3
Polymyxin B (300 units) Polymyxin B
Cell Membrane Disruption
17.6 ± 0.3 17.8 ± 0.4
Nalidixic acid (30 μg)
21.6 ± 0.3 20.4 ± 0.3
Ciprofloxacin(5 μg) Quinolones and
Fluoroquinolones
Nucleic Acid Synthesis inhibition
21.8 ± 0.2 22.0 ± 0.7
Chloramphenicol (30 μg) Chloramphenicol
9.8 ± 0.1 11.0 ± 0.4
Erythromycin (15 μg) Macrolides
24.0 ± 0 22.0 ± 0.1
Tetracyclin (30 μg) Tetracyclines
6.0 ± 0 6.0 ± 0
Novobiocin (5 μg)
10.8 ± 0.4 9.8 ± 0.1
Kanamycin (30 μg) Aminoglycosides
Protein Synthesis inhibition
13.4 ± 0.3 12.6 ± 0.2
Fosfomycin (200 μg) Fosfomycin
20.0 ± 1.0 18.0 ± 1.0
Ceftazidime (30 mcg)
23.0 ± 0.5 23.0 ± 0.9
Cefotaxime (30 μg) Cephalosporins
6.0 ± 0 6.0 ± 0
Penicillin G (10 units)
6.0 ± 0 6.0 ± 0
Ampicillin (10 μg) Penicillins
Cell Wall Synthesis Inhibition
rstB－ Wild type
Klebsiella pneumoniae CG43S3
23.2 ± 0.3 22.8 ± 0.2
Cefmetazole (30 μg)
6.0 ± 0 6.0 ± 0
Methicillin (5 mcg)
18.0 ± 0.2 17.0 ± 0.1
Carbenicillin (100 μg)
6.0 ± 0 6.0 ± 0
Sulfamethoxazole (25 μg) Sulfonamides
Antimetabolites
13.8 ± 0.4 13.6 ± 0.3
Polymyxin B (300 units) Polymyxin B
Cell Membrane Disruption
17.6 ± 0.3 17.8 ± 0.4
Nalidixic acid (30 μg)
21.6 ± 0.3 20.4 ± 0.3
Ciprofloxacin(5 μg) Quinolones and
Fluoroquinolones
Nucleic Acid Synthesis inhibition
21.8 ± 0.2 22.0 ± 0.7
Chloramphenicol (30 μg) Chloramphenicol
9.8 ± 0.1 11.0 ± 0.4
Erythromycin (15 μg) Macrolides
24.0 ± 0 22.0 ± 0.1
Tetracyclin (30 μg) Tetracyclines
6.0 ± 0 6.0 ± 0
Novobiocin (5 μg)
10.8 ± 0.4 9.8 ± 0.1
Kanamycin (30 μg) Aminoglycosides
Protein Synthesis inhibition
13.4 ± 0.3 12.6 ± 0.2
Fosfomycin (200 μg) Fosfomycin
20.0 ± 1.0 18.0 ± 1.0
Ceftazidime (30 mcg)
23.0 ± 0.5 23.0 ± 0.9
Cefotaxime (30 μg) Cephalosporins
6.0 ± 0 6.0 ± 0
Penicillin G (10 units)
6.0 ± 0 6.0 ± 0
Ampicillin (10 μg) Penicillins
Cell Wall Synthesis Inhibition
rstB－ Wild type
Klebsiella pneumoniae CG43S3
圖四. 分析克雷白氏肺炎桿菌中 RstA（A）或 RstB（B）對於不同藥物濾紙 錠的影響。將野生株、缺損株或帶有表現蛋白質質體的菌隔夜培養後，以 1:1 的比例和 LB 混合，於 37℃培養三十分鐘後，以棉花棒沾取菌液塗盤，
再放入不同藥物的濾紙錠，經隔夜培養後觀察抑菌圈直徑的大小。單位：
mm。pRK415 （vector only）；pHY110（containing putative rstA promoter region and rstA）；pHY112 （containing putative rstA promoter region, rstA, putative rstB promoter region, and rstB）。
8.6 ± 0 7.8 ± 0
8.8 ± 0 8.0 ± 0
Sulfamethoxazde (25 μg)
19.0 ± 1.5 6.0 ± 0
18.0 ± 1.0 6.0 ± 0
Sulfamethoxazole (25 μg) Sulfonamides
Polymyxin B (300 units) Polymyxin B
Cell Membrane Disruption
8.0 ± 0 6.0 ± 0
12.0 ± 1.0 6.0 ± 0
Nalidixic acid (30 μg)
30.6 ± 0.5 18.6 ± 0.5
32.0 ± 1.7 22.0 ± 0
Ciprofloxacin (5 μg) Quinolones and
Fluoroquinolones
Nucleic Acid Synthesis inhibition
19.6 ± 0.2 18.2 ± 0.2
24.0 ± 1.0 16.6 ± 0.5
Chloramphenicol (30 μg) Chloramphenicol
6.0 ± 0 6.0 ± 0
6.0 ± 0 6.0 ± 0
Erythromycin (15 μg) Macrolides
26.6 ± 0.5 19.2 ± 0.5
28.6 ± 0.5 20.5 ± 0.5
Tetracyclin (30 μg) Tetracyclines
6.0 ± 0 6.0 ± 0
6.0 ± 0 6.0 ± 0
Novobiocin (5 μg)
6.0 ± 0
Protein Synthesis inhibition
57.2 ± 0.5 50.2 ± 0.1
51.2 ± 1.1 52.4 ± 0.5
Fosfomycin (200 μg) Fosfomycin
33.2 ± 0.5 27.2 ± 0.5
38.0 ± 0 26.0 ± 1.0
Ceftazidime (30 mcg)
36.6 ± 0.5 26.6 ± 0.5
45.2 ± 1.5 30.0 ± 1.0
Cefotaxime (30 μg) Cephalosporins
10.0 ± 0 9.4 ± 0.2
11.0 ± 0.5 16.0 ± 0
Penicillin G (10 units)
23.6 ± 0.2 18.0 ± 1.0
27.2 ± 0.5 19.0 ± 0.5
Ampicillin (10 μg) Penicillins
Cell Wall Synthesis Inhibition
0.5 mM, 4h
E. coli BL21
Drug
8.6 ± 0 7.8 ± 0
8.8 ± 0 8.0 ± 0
Sulfamethoxazde (25 μg)
19.0 ± 1.5 6.0 ± 0
18.0 ± 1.0 6.0 ± 0
Sulfamethoxazole (25 μg) Sulfonamides
Polymyxin B (300 units) Polymyxin B
Cell Membrane Disruption
8.0 ± 0 6.0 ± 0
12.0 ± 1.0 6.0 ± 0
Nalidixic acid (30 μg)
30.6 ± 0.5 18.6 ± 0.5
32.0 ± 1.7 22.0 ± 0
Ciprofloxacin (5 μg) Quinolones and
Fluoroquinolones
Nucleic Acid Synthesis inhibition
19.6 ± 0.2 18.2 ± 0.2
24.0 ± 1.0 16.6 ± 0.5
Chloramphenicol (30 μg) Chloramphenicol
6.0 ± 0 6.0 ± 0
6.0 ± 0 6.0 ± 0
Erythromycin (15 μg) Macrolides
26.6 ± 0.5 19.2 ± 0.5
28.6 ± 0.5 20.5 ± 0.5
Tetracyclin (30 μg) Tetracyclines
6.0 ± 0 6.0 ± 0
6.0 ± 0 6.0 ± 0
Novobiocin (5 μg)
6.0 ± 0
Protein Synthesis inhibition
57.2 ± 0.5 50.2 ± 0.1
51.2 ± 1.1 52.4 ± 0.5
Fosfomycin (200 μg) Fosfomycin
33.2 ± 0.5 27.2 ± 0.5
38.0 ± 0 26.0 ± 1.0
Ceftazidime (30 mcg)
36.6 ± 0.5 26.6 ± 0.5
45.2 ± 1.5 30.0 ± 1.0
Cefotaxime (30 μg) Cephalosporins
10.0 ± 0 9.4 ± 0.2
11.0 ± 0.5 16.0 ± 0
Penicillin G (10 units)
23.6 ± 0.2 18.0 ± 1.0
27.2 ± 0.5 19.0 ± 0.5
Ampicillin (10 μg) Penicillins
Cell Wall Synthesis Inhibition
0.5 mM, 4h
E. coli BL21
Drug
圖五. 大腸桿菌中大量表現 RstA 對於不同藥物濾紙錠的影響。將帶有完整表 現 RstA（pHY089）或僅含 RstA N 端部分（pHY091）的大腸桿菌 BL21
（DE3）經隔夜培養後，取二十分之ㄧ加至 4 毫升的 LB 中，於 37℃培養至 OD600為 0.4~0.5，加入 0.5 mM IPTG 誘導三小時後，以棉花棒沾取菌液塗抹 於 LB 培養皿，再放入不同藥物的濾紙錠，經隔夜培養後，觀察抑菌圈直徑 的大小。單位：mm。
（A）
E. coli K-12 MG1655
asr
K. pneumoniae MGH 78578
asr
S. enterica serovar Typhimurium CT18
tnpA asr STY1579
STY1578 STY1576
bioD STY1583 STY1584
1 kb E. coli K-12 MG1655
asr
K. pneumoniae MGH 78578
asr
S. enterica serovar Typhimurium CT18
tnpA asr STY1579
STY1578 STY1576
bioD STY1583 STY1584
E. coli K-12 MG1655
asr
K. pneumoniae MGH 78578
asr
S. enterica serovar Typhimurium CT18
tnpA asr STY1579
STY1578 STY1576
bioD STY1583 STY1584
1 kb
（B）
cttgtctaagcccgccgctccggcgggctttttcgttcttcgctataacggcgcgttgTACATACAttccgtTACTTACTc
cctgacaccaaagactcactgatagcctcccggcaggcgcttatagtgttgccaacagccccgcagtggg
gttactcaagaaaccaagtcgaggatttcagaatg asr +1
-35 box -10 box putative RstA binding box cttgtctaagcccgccgctccggcgggctttttcgttcttcgctataacggcgcgttgTACATACAttccgtTACTTACTc
cctgacaccaaagactcactgatagcctcccggcaggcgcttatagtgttgccaacagccccgcagtggg
gttactcaagaaaccaagtcgaggatttcagaatg asr +1
-35 box -10 box putative RstA binding box
圖六. 分析不同菌種中 asr 上下游基因以及其可能的啟動子序列：（A）分析 大腸桿菌、克雷白氏肺炎桿菌、沙門氏菌的 asr 基因構造。基因說明如下：
ynfK : putative dethiobiotin synthetase, dgsA : DNA-binding transcriptional
repressor, ynfL : transcriptional regulator, ynfM : Inner membrane transport protein YnfM, ydgU : hypothetical protein, ydgD : predicted peptidase, mdtI : Multidrug resistance protein, mdt : multidrug efflux system protein MdtJ, ydhC : inner membrane transport protein YdhC, cfa : cyclopropane-fatty-acyl-phospholipid synthase family protein, ribC : riboflavin synthase alpha subunit, norM : multidrug efflux protein, KPN 02003 & KPN 02004 : hypothetical protein, bioD : putative dithiobiotin synthetase, STY 1576 & STY 1578 : putative regulatory protein, STY 1579 : putative membrane transport protein, tnpA : transposase for insertion
sequence element IS200, STY 1583 : putative secreted protein, STY 1584 : multidrug efflux system protein MdtI.（B）克雷白氏肺炎桿菌中，asr 可能的啟 動子區域（約 175 bp）之序列分析，灰底標示為可能的 RstA 鍵結辨認區域 (29)。
（A）
asr KPN 02004 norM
asr KPN 02004 norM
lacZ
1kb
pLacZ15
（B） （C）
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
（A） （B）
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
hour
（A） （B）
survival rate (%)
0
survival rate (%)
0
survival rate (%)
0
survival rate (%)
0
（A）
（B）
asr KPN 02004 norM
KPN 02003 768 bp
Deletion
YWp16 294 bp
YWp17 asr KPN 02004 norM
KPN 02003 768 bp
Deletion
YWp16 294 bp
YWp17
wild type form mutant form
wild type form mutant form
Loss of plasmid
△asr
Kp CG43S3
△asr
△asr in pKAS46 asr Homologous recombination
K. pneumoniae CG43S3
asr
Delete, rejoin and clone into a suicide vector
Loss of plasmid
△asr
Kp CG43S3
△asr
△asr in pKAS46 asr Homologous recombination
K. pneumoniae CG43S3
asr
Delete, rejoin and clone into a suicide vector
（A）
survival rate (%)
0
wild type △asr adapted wild type △asr
unadapted
survival rate (%)
0
wild type △asr adapted wild type △asr
unadapted
survival rate (%)
0
survival rate (%)
0
圖十二. asr 的基因缺損在酸環境下的生長曲線及酸逆境下的存活率。（A）
取二十分之ㄧ隔夜培養的菌液加入 pH 7.0、pH 4.8、pH 4.6、pH 4.4 LPM 培 養液中，於不同時間點偵測 600 nm 的吸光值。（B）野生株及 asr 缺損株於 酸逆境下的存活率，測試與計數方式同圖十 A。（C）asr 互補株在酸逆境下 的存活：將轉殖帶有質體的細菌株培養至 OD600＝0.6 後，塗盤數菌，接著弱 酸 pH 4.6 LPM 培養液適應兩小時後再給予 pH 3.0 LPM 培養液酸逆境兩小 時，再塗盤，隔夜計數酸逆境後塗盤的菌數除以初始塗盤菌數為存活率。
（A）
圖十三. 重組蛋白 Asr 的表現純化及西方墨點法分析。 （A）大腸桿菌 BL21
（DE3）[ pGEX-5X-1-Asr ]以不同濃度 IPTG 誘導：lanes 1 和 12 未添加 IPTG； lanes 3、4、5，添加 0.1 mM IPTG； lanes 2、6、7、8，添加 0.5 mM IPTG；lanes 9、10、11，添加 IPTG 1 mM。IPTG 誘導表現時間分別為兩小 時（lanes 3、6、9）、四小時（lanes 4、7、10）、六小時（lanes 1、2、5、
8、11、12）後以 SDS-PAGE 分析。（B）膠片以 Coomassie Blue 染色或
（C）GST 抗體做免疫呈色。Lane 1：分子量標記；2： BL21（DE3）
[pGEX-5X-1]以 0.5 mM IPTG 誘導六小時；3：BL21（DE3）[pGEX-5X-1-Asr]
未加 IPTG 培養六小時； 4：BL21（DE3）[pGEX-5X-1-Asr]以 0.5 mM IPTG 誘導六小時； 5：BL21（DE3）[pGEX-5X-1-Asr]以 0.5 mM IPTG 誘導六小時 後，經超音波震盪所離心的沉澱物； 6：BL21（DE3）[pGEX-5X-1-Asr]以 0.5 mM IPTG 誘導六小時，經超音波震盪並離心後的上清液；7：經 GST 管 柱純化後的重組蛋白Asr。
（A） （B） 動子的活性表現。pHY112 （containing putative rstA promoter region, rstA, putative rstB promoter region, and rstB）
Z01 Z01△rstA
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
Z01 Z01△rstA Z01 Z01△rstA Z01 Z01△rstA FeSO440 µM 2,2-dipyridyl 0.2 mM
β−galactosidase activity (Miller units)
0
Z01 Z01△rstA Z01 Z01△rstA Z01 Z01△rstA FeSO440 µM 2,2-dipyridyl 0.2 mM
K. pneumoniae CG43S3
E. coli K-12 MG1655
S. Typhimurium LT2
GTTAGGGCAGCATAGCGGATTTAACGCTGTTTATAAAAGAGTAATGGCGGGT AAAATAAGTGCGGTGTATGTTACTTCTTTTACTGATGTGCAATAGAC-ATG
-35 -10
GTCGGGAAAAGTGGAATCAGCCCGGCGATATAATAATTTTTCGTTTTTGCTAAA ACACCAATCAACAGCACTACCAGCGCACCGAGCGCGGCTTTGATTACCAGCC CCATCTTTTTACCTTAACACTTCCATAACAAGTCATCAGTAGAATACCTGATGAA AACTTGTTTAGAAACGATTGATAGTAAGTAAAAACAGCGCG-GTG
-35 -10
ACGTTGGAAACAGAGGAATTAATCCGGCGATATAATAATTTTTTGTTTTTGACAG CAGACCAATCAATACGACGACCAGCGCCCCCAGGGCAGCTTTAATCACGAGTC CCATTACCTTGCCTTAACATGCTAATAACAACAGCATGTAGCATAACGGAACCG CTCTCGTTTAGAAAAGATTTATG-53bp-ATG
-35 -10
K. pneumoniae CG43S3
E. coli K-12 MG1655
S. Typhimurium LT2
GTTAGGGCAGCATAGCGGATTTAACGCTGTTTATAAAAGAGTAATGGCGGGT AAAATAAGTGCGGTGTATGTTACTTCTTTTACTGATGTGCAATAGAC-ATG
-35 -10
GTCGGGAAAAGTGGAATCAGCCCGGCGATATAATAATTTTTCGTTTTTGCTAAA ACACCAATCAACAGCACTACCAGCGCACCGAGCGCGGCTTTGATTACCAGCC CCATCTTTTTACCTTAACACTTCCATAACAAGTCATCAGTAGAATACCTGATGAA AACTTGTTTAGAAACGATTGATAGTAAGTAAAAACAGCGCG-GTG
-35 -10
ACGTTGGAAACAGAGGAATTAATCCGGCGATATAATAATTTTTTGTTTTTGACAG CAGACCAATCAATACGACGACCAGCGCCCCCAGGGCAGCTTTAATCACGAGTC CCATTACCTTGCCTTAACATGCTAATAACAACAGCATGTAGCATAACGGAACCG CTCTCGTTTAGAAAAGATTTATG-53bp-ATG
-35 -10
PhoP consensus sequence (T/G)GTTTA-nnnnn-(T/G)GTTTA RstA consensus sequence TACA-nnnnnn-TACA
PhoP consensus sequence (T/G)GTTTA-nnnnn-(T/G)GTTTA RstA consensus sequence TACA-nnnnnn-TACA
圖十五. rstA 啟動子區域序列分析。克雷白氏肺炎桿菌、大腸桿菌、以及沙 門氏菌的 rstA 啟動子區域，以方框標示 PhoP 辨認位置 (25)，以虛線標示 RstA 辨認序列 (29)。
0 20 40 60 80 100 120
low Mg2+(-) high Mg2+(MgCl
2) high Mg2+(MgSO4)
β−galactosidase activity (Miller units)
Z01 Z01△phoP 0
20 40 60 80 100 120
low Mg2+(-) high Mg2+(MgCl
2) high Mg2+(MgSO4)
β−galactosidase activity (Miller units)
Z01 Z01△phoP
圖十六. PhoP 對於 rstA 啟動子活性的分析。取二十分之ㄧ隔夜培養的菌液加 至4 毫升新鮮的 LB 培養液中，分別未添加或加入 30 mM氯化鎂或硫酸鎂，
37℃培養至 OD600=0.6，依據 Miller 的方法操作，偵測 rstA 啟動子的活性表 現。
β−galactosidase activity (Miller units) 0 20 40 60 80 100
low Mg2+ (-) high Mg2+ (MgCl2) high Mg2+ (MgSO4)
Z01 Z01△phoP
β−galactosidase activity (Miller units)
0 20 40 60 80 100
low Mg2+ (-) high Mg2+ (MgCl2) high Mg2+ (MgSO4)
Z01 Z01△phoP
圖十七. PhoP 對於 rstB 啟動子活性的分析。取二十分之ㄧ隔夜培養的菌液加 至4 毫升的 LPM 培養液中，分別未添加或加入 30 mM 氯化鎂或硫酸鎂，37
℃培養至OD600=0.6，依據 Miller 的方法操作，偵測 rstB 啟動子的活性表 現。
β−galactosidase activity (Miller units)
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
（A） （B）
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0 OD600=0.5，分別測 rstA、rstB、asr 啟動子的活性表現。加入的金屬離子分別 為1 mM CaCl2、1 mM CuSO4、30 mM MgCl2、1 mM ZnCl2、1 mM FeCl3、 30 mM MgSO 、1 mM FeSO 。
（A）
-8 tgtt gataat:gggaat:ctttatc gaca +19 iucA -82 aaac acaaat:gataat:cattatc atct -55 iroB -138 tatt gatgat:aaaaac:cattctc atta -111 feoA -89 gttc tataat:gagacg:cattacg tcgg -62 fur
.atc GATAAT:GATAAT:CATTATC tac. Fur Box -87 cggg gatatc:gataa c:attgact cccc -60 rstB -8 tgtt gataat:gggaat:ctttatc gaca +19 iucA -82 aaac acaaat:gataat:cattatc atct -55 iroB -138 tatt gatgat:aaaaac:cattctc atta -111 feoA -89 gttc tataat:gagacg:cattacg tcgg -62 fur
.atc GATAAT:GATAAT:CATTATC tac. Fur Box -87 cggg gatatc:gataa c:attgact cccc -60 rstB
（B）
LPM pH 7.0 LPM pH 7.0 LPM pH 4.6
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0
galactosidase activity (Miller units)
0
β−galactosidase activity (Miller units)
0