Nitric Oxide Inhibitory Meroterpenoids from the Fungus Penicillium ...
Nitric Oxide Inhibitory Meroterpenoids from the Fungus Penicillium purpurogenum MHZ 111 Jing Sun,†,‡ Zhi-Xiang Zhu,† Yue-Lin Song,† Dan Dong,§ Jiao Zheng,† Ting Liu,§ Yun-Fang Zhao,† Daneel Ferreira,┴ Jordan K. Zjawiony,┴ Peng-Fei Tu,*,† and Jun Li*,†
†
Modern Research Center for Traditional Chinese Medicine, Beijing University of
Chinese Medicine, Beijing 100029, People’s Republic of China ‡
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing
100102, People’s Republic of China §
Institute of Plant and Environment Protection, Beijing Academy of Agriculture and
Forestry Sciences, Beijing 100097, People’s Republic of China ┴
Department of BioMolecular Sciences, Division of Pharmacognosy, and Research
Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, University, Mississippi 38677-1848, United States
1
The list of contents No.
Content
S1
1
S2
13
S3
gHSQC spectrum of purpurogenolide A (1)
S4
gHMBC spectrum of purpurogenolide A (1)
S5
NOESY spectrum of purpurogenolide A (1)
S6
HRESIMS spectrum of purpurogenolide A (1)
S7
1
S8
13
S9
gHSQC spectrum of purpurogenolide B (2)
S10
gHMBC spectrum of purpurogenolide B (2)
S11
NOESY spectrum of purpurogenolide B (2)
S12
HRESIMS spectrum of purpurogenolide B (2)
S13
DFT-optimized structures for low-energy conformers of
H NMR spectrum of purpurogenolide A (1) C NMR spectrum of purpurogenolide A (1)
H NMR spectrum of purpurogenolide B (2) C NMR spectrum of purpurogenolide B (2)
(4R,5R,7R,9S,11R,12S,22R)-2 S14
1
S15
13
S16
gHSQC spectrum of purpurogenolide C (3)
S17
gHMBC spectrum of purpurogenolide C (3)
S18
NOESY spectrum of purpurogenolide C (3)
S19
HRESIMS spectrum of purpurogenolide C (3)
S20
DFT-optimized structures for low-energy conformers of
H NMR spectrum of purpurogenolide C (3) C NMR spectrum of purpurogenolide C (3)
(4R,5R,7R,9S,11R,12S,22R)-3 S21
1
S22
13
S23
gHSQC spectrum of purpurogenolide C (4)
S24
gHMBC spectrum of purpurogenolide D (4)
S25
NOESY spectrum of purpurogenolide D (4)
H NMR spectrum of purpurogenolide D (4) C NMR spectrum of purpurogenolide D (4)
2
S26
HRESIMS spectrum of purpurogenolide D (4)
S27
DFT-optimized structures for low-energy conformers of (4S,5S,7S,9S,10R,11R,12R,13R,22S,23S)-4
S28
1
S29
13
S30
gHSQC spectrum of purpurogenolide E (5)
S31
gHMBC spectrum of purpurogenolide E (5)
S32
NOESY spectrum of purpurogenolide E (5)
S33
Selected NOESY correlations of purpurogenolide E (5)
S34
HRESIMS spectrum of purpurogenolide E (5)
S35
The morphology of Penicillium purpurogenum MHZ111.
S36
The 18S rDNA sequences data of Penicillium purpurogenum MHZ 111.
S37
The ITS rDNA sequences data of Penicillium purpurogenum MHZ 111.
H NMR spectrum of purpurogenolide E (5) C NMR spectrum of purpurogenolide E (5)
3
O O
H O O
O
O
O
S1 1H NMR spectrum of purpurogenolide A (1) (500 MHz, methanol-d4)
O O
H O O
O
O
O
S2 13C NMR spectrum of purpurogenolide A (1) (125 MHz, methanol-d4) 4
O O
H O O
O
O
O
S3 gHSQC spectrum of purpurogenolide A (1) (500 MHz, methanol-d4) O O
H O O
O
O
O
S4 gHMBC spectrum of purpurogenolide A (1) (500 MHz, methanol-d4) 5
O O
H O O
O
O
O
S5 NOESY spectrum of purpurogenolide A (1) (500 MHz, methanol-d4)
6
O O
H O O
O O
S6 HRESIMS spectrum of purpurogenolide A (1)
7
O
O HO
H
OH
O
O
O
O
O O
S7 1H NMR spectrum of purpurogenolide B (2) (500 MHz, CDCl3)
O HO
H
O
O
OH O
O
O O
S8 13C NMR spectrum of purpurogenolide B (2) (125 MHz, CDCl3) 8
O HO
H
OH
O
O
O
O
O O
S9 HSQC spectrum of purpurogenolide B (2) (500 MHz, CDCl3) O HO
H
O
O
OH O
O
O O
S10 HMBC spectrum of purpurogenolide B (2) (500 MHz, CDCl3) 9
O HO
H
O
O
OH O
O
O O
S11 NOESY spectrum of purpurogenolide B (2) (500 MHz, CDCl3)
10
O HO
H
O
O
OH O
O
O O
S12 HRESIMS spectrum of purpurogenolide B (2)
2C1 (100%) S13 DFT-optimized structures for low-energy conformers of (4R,5R,7R,9S,11R,12S,22R)-2 11
O H O
O H
H O
O O
O O
S14 1H NMR spectrum of purpurogenolide C (3) (500 MHz, methanol-d4)
O H O
O H
H O
O O
O O
S15 13C NMR spectrum of purpurogenolide C (3) (125 MHz, methanol-d4) 12
O H
O
O
H
H O
O O
O O
S16 gHSQC spectrum of purpurogenolide C (3) (500 MHz, methanol-d4) O H O
O H
H O
O O
O O
S17 gHMBC spectrum of purpurogenolide C (3) (500 MHz, methanol-d4) 13
O H O
O H
H O
O O
O O
S18 NOESY spectrum of purpurogenolide C (3) (500 MHz, methanol-d4)
14
O H O
O H
O O
O O
S19 HRESIMS spectrum of purpurogenolide C (3)
3C1 (0.031%)
3C2 (0.29%)
3C3 (99.67%)
S20 DFT-optimized structures for low-energy conformers of (4R,5R,7R,9S,11R,12S,22R)-3 15
H O
O OH O
O H
H O
O O 17
H3CO
OH O
S21 1H NMR spectrum of purpurogenolide D (4) (500 MHz, methanol-d4)
O OH O
O H
H O
O O 17
H3CO
OH O
S22 13C NMR spectrum of purpurogenolide D (4) (125 MHz, methanol-d4)
16
O OH O
O H
H O
O O 17
H3CO
OH O
S23 gHSQC spectrum of purpurogenolide D (4) (500 MHz, methanol-d4) O OH O
O H
H O
O O 17
H3CO
OH O
S24 gHMBC spectrum of purpurogenolide D (4) (500 MHz, methanol-d4) 17
O OH O
O H
H O
O O 17
H3CO
OH O
S25 NOESY spectrum of purpurogenolide D (4) (500 MHz, methanol-d4)
18
O OH O
O H
H O
O O 17
H3CO
OH O
S26 HRESIMS spectrum of purpurogenolide D (4)
4C1 (49.95%)
4C2 (50.05%)
S27 DFT-optimized structures for low-energy conformers of (4S,5S,7S,9S,10R,11R,12R,13R,22S,23S)-4 19
O
OH
O O
H O
H O
H OO
S28 1H NMR spectrum of purpurogenolide E (5) (500 MHz, methanol-d4)
O
OH
O O
H O
H O
H OO
S29 13C NMR spectrum of purpurogenolide E (5) (125 MHz, methanol-d4) 20
O
OH
O O
H O
H O
H OO
S30 gHSQC spectrum of purpurogenolide E (5) (500 MHz, methanol-d4) O
OH
O O
H O
H O
H OO
S31 gHMBC spectrum of purpurogenolide E (5) (500 MHz, methanol-d4) 21
O
OH
O O
H O
H O
H OO
S32 NOESY spectrum of purpurogenolide E (5) (500 MHz, methanol-d4)
S33 Selected NOESY correlations of 5.
22
O
OH
O O
H O
H O
H OO
S34 HRESIMS spectrum of purpurogenolide E (5)
A
B
20 μm
1 cm
S35 The morphology of Penicillium purpurogenum MHZ111. (A) Colony on Martin medium; (B) Conidiophore peduncle under light microscope. 23
S36 The 18S rDNA sequences data of Penicillium purpurogenum MHZ 111. CCAGTAGTCATATGCTTGTCTCAAAGATTAAGCCATGCATGTCTAAGTATAA GCACTCTTTTACTGTGAAACTGCGAATGGCTCATTAAATCAGTTATCGTTTA TTTGATAGTACCCTACTACATGGATACCTGTGGTAATTCTAGAGCTAATACAT GCGCAAAACCCCGACTTCGGAAGGGGTGTATTTATTAGATAAAAAACCAAT GCCCTTCGGGGCTCCTTGGTGATTCATAATAACTTCACGAATCGCATGGCCT TGCGCCGGCGATGGTTCATTCAAATTTCTGCCCTATCAACTTTCGATGGTAG GATAGTGGCCTACCATGGTGGCAACGGGTAACGGGGAATTAGGGTTCGATT CCGGAGAGGGAGCCTGAGAAACGGCTACCACATCCAAGGAAGGCAGCAG GCGCGCAAATTACCCAATCCCGATACGGGGAGGTAGTGACAATAAATACTG ATACAGGGCTCTTTTGGGTCTTGTAATTGGAATGAGAACAATCTAAATCCCT TAACGAGGAACAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATT CCAGCTCCAATAGCGTATATTAAAGTTGTTGCAGTTAAAAAGCTCGTAGTTG AACCTTGGGCCCGTCCTGCCGGTCCGCCTCACCGCGAGTACTGGTCCGGAT GGGCCTTTCTTTCTGGGGAATCCCATGGCCTTCACTGGCTGTGGCGGGGAA CCAGGACTTTTACTGTGAAAAAATTAGAGTGTTCAAAGCAGGCCTTTGCTC GGATACATTAGCATGGAATAATAGAATAGGACGTGCGGTTCTATTTTGTTGG TTTCTAGGACCGCCGTAATGATTAATAGGGATAGTCGGGGGCGTCAGTATTC AGCTGTCAGAGGTGAAATTCTTGGATTTGCTGAAGACTAACTACTGCGAAA GCATTCGCCAAGGATGTTTTCATTAATCAGGGAACGAAAGTTAGGGGATCG AAGACGATCAGATACCGTCGTAGTCTTAACCATAAACTATGCCGACTAGGG ATCGGGCGGGGTTTCTATGATGACCCGCTCGGCACCTTACGAGAAATCAAA 24
GTTTTTGGGTTCTGGGGGGAGTATGGTCGCAAGGCTGAAACTTAAAGAAAT TGACGGAAGGGCACCACAAGGCGTGGAGCCTGCGGCTTAATTTGACTCAA CACGGGGAAACTCACCAGGTCCAGACAAAATAAGGATTGACAGATTGAGA GCTCTTTCTTGATCTTTTGGATGGTGGTGCATGGCCGTTCTTAGTTGGTGGA GTGATTTGTCTGCTTAATTGCGATAACGAACGAGACCTCGGCCCTTAAATAG CCCGGTCCGCGTTTGCGGGCCGCTGGCTTCTTAGGGGGACTATCGGCTCAA GCCGATGGAAGTACGTGGCAATAACAGGTCTGTGATGCCCTTAGATGTTCT GGGCCGCACGCGCGCTACACTGACAGGGCCAGCGAGTACATCACCTTGGC CGAGAGGTCTGGGTAATCTTGTTAAACCCTGTCGTGCTGGGGATAGAGCAT TGCAATTATTGCTCTTCAACGAGGAATGCCTAGTAGGCACGAGTCATCAGC TCGTGCCGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTACTACCG ATTGAATGGCT
S37 The ITS rDNA sequences data of Penicillium purpurogenum MHZ 111. TCCGTAGGTGAACCTGCGGAAGGATCATTACCGAGTGCGGGCCCTCGTGGC CCAACCTCCCACCCTTGTCTCTATACACCTGTTGCTTTGGCGGGCCCACCG GGGCCACCTGGTCGCCGGGGGACATCTGTCCCCGGGCCCGCGCCCGCCGA AGCGCTCTGTGAACCCTGATGAAGATGGGCTGTCTGAGTACTATGAAAATT GTCAAAACTTTCAACAATGGATCTCTTGGTTCCGGCATCGATGAAGAACGC AGCA
25
†
Modern Research Center for Traditional Chinese Medicine, Beijing University of
Chinese Medicine, Beijing 100029, People’s Republic of China ‡
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing
100102, People’s Republic of China §
Institute of Plant and Environment Protection, Beijing Academy of Agriculture and
Forestry Sciences, Beijing 100097, People’s Republic of China ┴
Department of BioMolecular Sciences, Division of Pharmacognosy, and Research
Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, University, Mississippi 38677-1848, United States
1
The list of contents No.
Content
S1
1
S2
13
S3
gHSQC spectrum of purpurogenolide A (1)
S4
gHMBC spectrum of purpurogenolide A (1)
S5
NOESY spectrum of purpurogenolide A (1)
S6
HRESIMS spectrum of purpurogenolide A (1)
S7
1
S8
13
S9
gHSQC spectrum of purpurogenolide B (2)
S10
gHMBC spectrum of purpurogenolide B (2)
S11
NOESY spectrum of purpurogenolide B (2)
S12
HRESIMS spectrum of purpurogenolide B (2)
S13
DFT-optimized structures for low-energy conformers of
H NMR spectrum of purpurogenolide A (1) C NMR spectrum of purpurogenolide A (1)
H NMR spectrum of purpurogenolide B (2) C NMR spectrum of purpurogenolide B (2)
(4R,5R,7R,9S,11R,12S,22R)-2 S14
1
S15
13
S16
gHSQC spectrum of purpurogenolide C (3)
S17
gHMBC spectrum of purpurogenolide C (3)
S18
NOESY spectrum of purpurogenolide C (3)
S19
HRESIMS spectrum of purpurogenolide C (3)
S20
DFT-optimized structures for low-energy conformers of
H NMR spectrum of purpurogenolide C (3) C NMR spectrum of purpurogenolide C (3)
(4R,5R,7R,9S,11R,12S,22R)-3 S21
1
S22
13
S23
gHSQC spectrum of purpurogenolide C (4)
S24
gHMBC spectrum of purpurogenolide D (4)
S25
NOESY spectrum of purpurogenolide D (4)
H NMR spectrum of purpurogenolide D (4) C NMR spectrum of purpurogenolide D (4)
2
S26
HRESIMS spectrum of purpurogenolide D (4)
S27
DFT-optimized structures for low-energy conformers of (4S,5S,7S,9S,10R,11R,12R,13R,22S,23S)-4
S28
1
S29
13
S30
gHSQC spectrum of purpurogenolide E (5)
S31
gHMBC spectrum of purpurogenolide E (5)
S32
NOESY spectrum of purpurogenolide E (5)
S33
Selected NOESY correlations of purpurogenolide E (5)
S34
HRESIMS spectrum of purpurogenolide E (5)
S35
The morphology of Penicillium purpurogenum MHZ111.
S36
The 18S rDNA sequences data of Penicillium purpurogenum MHZ 111.
S37
The ITS rDNA sequences data of Penicillium purpurogenum MHZ 111.
H NMR spectrum of purpurogenolide E (5) C NMR spectrum of purpurogenolide E (5)
3
O O
H O O
O
O
O
S1 1H NMR spectrum of purpurogenolide A (1) (500 MHz, methanol-d4)
O O
H O O
O
O
O
S2 13C NMR spectrum of purpurogenolide A (1) (125 MHz, methanol-d4) 4
O O
H O O
O
O
O
S3 gHSQC spectrum of purpurogenolide A (1) (500 MHz, methanol-d4) O O
H O O
O
O
O
S4 gHMBC spectrum of purpurogenolide A (1) (500 MHz, methanol-d4) 5
O O
H O O
O
O
O
S5 NOESY spectrum of purpurogenolide A (1) (500 MHz, methanol-d4)
6
O O
H O O
O O
S6 HRESIMS spectrum of purpurogenolide A (1)
7
O
O HO
H
OH
O
O
O
O
O O
S7 1H NMR spectrum of purpurogenolide B (2) (500 MHz, CDCl3)
O HO
H
O
O
OH O
O
O O
S8 13C NMR spectrum of purpurogenolide B (2) (125 MHz, CDCl3) 8
O HO
H
OH
O
O
O
O
O O
S9 HSQC spectrum of purpurogenolide B (2) (500 MHz, CDCl3) O HO
H
O
O
OH O
O
O O
S10 HMBC spectrum of purpurogenolide B (2) (500 MHz, CDCl3) 9
O HO
H
O
O
OH O
O
O O
S11 NOESY spectrum of purpurogenolide B (2) (500 MHz, CDCl3)
10
O HO
H
O
O
OH O
O
O O
S12 HRESIMS spectrum of purpurogenolide B (2)
2C1 (100%) S13 DFT-optimized structures for low-energy conformers of (4R,5R,7R,9S,11R,12S,22R)-2 11
O H O
O H
H O
O O
O O
S14 1H NMR spectrum of purpurogenolide C (3) (500 MHz, methanol-d4)
O H O
O H
H O
O O
O O
S15 13C NMR spectrum of purpurogenolide C (3) (125 MHz, methanol-d4) 12
O H
O
O
H
H O
O O
O O
S16 gHSQC spectrum of purpurogenolide C (3) (500 MHz, methanol-d4) O H O
O H
H O
O O
O O
S17 gHMBC spectrum of purpurogenolide C (3) (500 MHz, methanol-d4) 13
O H O
O H
H O
O O
O O
S18 NOESY spectrum of purpurogenolide C (3) (500 MHz, methanol-d4)
14
O H O
O H
O O
O O
S19 HRESIMS spectrum of purpurogenolide C (3)
3C1 (0.031%)
3C2 (0.29%)
3C3 (99.67%)
S20 DFT-optimized structures for low-energy conformers of (4R,5R,7R,9S,11R,12S,22R)-3 15
H O
O OH O
O H
H O
O O 17
H3CO
OH O
S21 1H NMR spectrum of purpurogenolide D (4) (500 MHz, methanol-d4)
O OH O
O H
H O
O O 17
H3CO
OH O
S22 13C NMR spectrum of purpurogenolide D (4) (125 MHz, methanol-d4)
16
O OH O
O H
H O
O O 17
H3CO
OH O
S23 gHSQC spectrum of purpurogenolide D (4) (500 MHz, methanol-d4) O OH O
O H
H O
O O 17
H3CO
OH O
S24 gHMBC spectrum of purpurogenolide D (4) (500 MHz, methanol-d4) 17
O OH O
O H
H O
O O 17
H3CO
OH O
S25 NOESY spectrum of purpurogenolide D (4) (500 MHz, methanol-d4)
18
O OH O
O H
H O
O O 17
H3CO
OH O
S26 HRESIMS spectrum of purpurogenolide D (4)
4C1 (49.95%)
4C2 (50.05%)
S27 DFT-optimized structures for low-energy conformers of (4S,5S,7S,9S,10R,11R,12R,13R,22S,23S)-4 19
O
OH
O O
H O
H O
H OO
S28 1H NMR spectrum of purpurogenolide E (5) (500 MHz, methanol-d4)
O
OH
O O
H O
H O
H OO
S29 13C NMR spectrum of purpurogenolide E (5) (125 MHz, methanol-d4) 20
O
OH
O O
H O
H O
H OO
S30 gHSQC spectrum of purpurogenolide E (5) (500 MHz, methanol-d4) O
OH
O O
H O
H O
H OO
S31 gHMBC spectrum of purpurogenolide E (5) (500 MHz, methanol-d4) 21
O
OH
O O
H O
H O
H OO
S32 NOESY spectrum of purpurogenolide E (5) (500 MHz, methanol-d4)
S33 Selected NOESY correlations of 5.
22
O
OH
O O
H O
H O
H OO
S34 HRESIMS spectrum of purpurogenolide E (5)
A
B
20 μm
1 cm
S35 The morphology of Penicillium purpurogenum MHZ111. (A) Colony on Martin medium; (B) Conidiophore peduncle under light microscope. 23
S36 The 18S rDNA sequences data of Penicillium purpurogenum MHZ 111. CCAGTAGTCATATGCTTGTCTCAAAGATTAAGCCATGCATGTCTAAGTATAA GCACTCTTTTACTGTGAAACTGCGAATGGCTCATTAAATCAGTTATCGTTTA TTTGATAGTACCCTACTACATGGATACCTGTGGTAATTCTAGAGCTAATACAT GCGCAAAACCCCGACTTCGGAAGGGGTGTATTTATTAGATAAAAAACCAAT GCCCTTCGGGGCTCCTTGGTGATTCATAATAACTTCACGAATCGCATGGCCT TGCGCCGGCGATGGTTCATTCAAATTTCTGCCCTATCAACTTTCGATGGTAG GATAGTGGCCTACCATGGTGGCAACGGGTAACGGGGAATTAGGGTTCGATT CCGGAGAGGGAGCCTGAGAAACGGCTACCACATCCAAGGAAGGCAGCAG GCGCGCAAATTACCCAATCCCGATACGGGGAGGTAGTGACAATAAATACTG ATACAGGGCTCTTTTGGGTCTTGTAATTGGAATGAGAACAATCTAAATCCCT TAACGAGGAACAATTGGAGGGCAAGTCTGGTGCCAGCAGCCGCGGTAATT CCAGCTCCAATAGCGTATATTAAAGTTGTTGCAGTTAAAAAGCTCGTAGTTG AACCTTGGGCCCGTCCTGCCGGTCCGCCTCACCGCGAGTACTGGTCCGGAT GGGCCTTTCTTTCTGGGGAATCCCATGGCCTTCACTGGCTGTGGCGGGGAA CCAGGACTTTTACTGTGAAAAAATTAGAGTGTTCAAAGCAGGCCTTTGCTC GGATACATTAGCATGGAATAATAGAATAGGACGTGCGGTTCTATTTTGTTGG TTTCTAGGACCGCCGTAATGATTAATAGGGATAGTCGGGGGCGTCAGTATTC AGCTGTCAGAGGTGAAATTCTTGGATTTGCTGAAGACTAACTACTGCGAAA GCATTCGCCAAGGATGTTTTCATTAATCAGGGAACGAAAGTTAGGGGATCG AAGACGATCAGATACCGTCGTAGTCTTAACCATAAACTATGCCGACTAGGG ATCGGGCGGGGTTTCTATGATGACCCGCTCGGCACCTTACGAGAAATCAAA 24
GTTTTTGGGTTCTGGGGGGAGTATGGTCGCAAGGCTGAAACTTAAAGAAAT TGACGGAAGGGCACCACAAGGCGTGGAGCCTGCGGCTTAATTTGACTCAA CACGGGGAAACTCACCAGGTCCAGACAAAATAAGGATTGACAGATTGAGA GCTCTTTCTTGATCTTTTGGATGGTGGTGCATGGCCGTTCTTAGTTGGTGGA GTGATTTGTCTGCTTAATTGCGATAACGAACGAGACCTCGGCCCTTAAATAG CCCGGTCCGCGTTTGCGGGCCGCTGGCTTCTTAGGGGGACTATCGGCTCAA GCCGATGGAAGTACGTGGCAATAACAGGTCTGTGATGCCCTTAGATGTTCT GGGCCGCACGCGCGCTACACTGACAGGGCCAGCGAGTACATCACCTTGGC CGAGAGGTCTGGGTAATCTTGTTAAACCCTGTCGTGCTGGGGATAGAGCAT TGCAATTATTGCTCTTCAACGAGGAATGCCTAGTAGGCACGAGTCATCAGC TCGTGCCGATTACGTCCCTGCCCTTTGTACACACCGCCCGTCGCTACTACCG ATTGAATGGCT
S37 The ITS rDNA sequences data of Penicillium purpurogenum MHZ 111. TCCGTAGGTGAACCTGCGGAAGGATCATTACCGAGTGCGGGCCCTCGTGGC CCAACCTCCCACCCTTGTCTCTATACACCTGTTGCTTTGGCGGGCCCACCG GGGCCACCTGGTCGCCGGGGGACATCTGTCCCCGGGCCCGCGCCCGCCGA AGCGCTCTGTGAACCCTGATGAAGATGGGCTGTCTGAGTACTATGAAAATT GTCAAAACTTTCAACAATGGATCTCTTGGTTCCGGCATCGATGAAGAACGC AGCA
25
