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*��35#2!�����,3B:(%�:�� */�1@3 �3 ��& '"���?0<� C��
�A �������B ������A �������B �����
P-25
�ßá2Å��Ù¦@XH8î"�ùÆÁÖ#�Á�)��pÅ�/��"�Û��.������"¢ã"�È� .ñjë�Ö!���#�_"��0GT5Y<RX#� ��ârk �!+.�²º!t�Õf�ÆÁuÀ"Ā�©� ¢ã"sl�®��/.�°ÍÐ�#�h|¡�ľ��.<3FG8BT3"h�Òñjq�¹¶2�,!«,�!�.��)!��"Ìå2pÅ���ßá!�.Ù¦@XH8î"ÆÁ2h�ë��.���� .�hÆÁ<=BN�"úÆ2ÉÇ����
Dark
!����
���#��
Light
����� ����������
� ������
�"���Promoter
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���#��
� ������
�"���Promoter
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(A)
'"9$�=6
� ����� NblS-RpaB8.;-)%
°ÍÐ�#�he�Ç ÆÁìÓ!û1.ñj~�����<3FG8BT3Synechococcus elongatus PCC7942"V=LX=V7QVY@YRpaB!ËÉ����/&�"ÍÐ��RpaB#�h=CV=�Ò!û1.���Ï�/��.āAĂ�ò�±a�#RpaB#TXøv�/�hliA �¸Çöb�[¼"HLR1þ�!Ú|�ñj2£q�.���h±a�#ó*�!ÞTXøv�/ñj�ú��.���,�öb�ÆÁë�Ö!�.g/�TJVA:Y���"pÅ�®��/������RpaB�»�ø× �h=CV=!�Ò�."���.�#hàð!�Ò�."�#]«�����(��· h���v!ý��ñjq�2éØ!µç��āB-DĂ�S. elongatus2·� h����ÿ����+-��h±a!<IC��ý"RpaB"TXøv¿ "�vāPhos-tag2Å��SDS-PAGEă{Ă�öb�ÆÁH@YXāF;Xæ³:�Ă2�/�/ì%���"Ú´��h"' ,��¬→«*�h→ò�h �`"h±a!��)hliA �"ÆÁ�ë��/��/!c��RpaB�ÞTXøv�/.���Ï�/��(�RpaB!+.ñjq�Ö#��h=CV=!ü,��· hÃ��v!���û^�.���Ï}�/��
RpaB�A?��*:(>7%+4
PhliAKpnI
PstIPstI
PcphIRpaB
SacII
PcphI CphI NblS
SmaIXmaI
……
Target gene
PcphI-CphI-NblS�Light Sensor�HK�PcphI-RpaB�RR�
� ��
PhliAKpnI
PstIPstI
PcphIRpaB
SacII
PcphI NblS
SmaIXmaI……
Target gene
PcphI-NblS�HK�PcphI-RpaB�RR�
� ��
Light
HLR1 motif
NblS
RpaB
High Light
P
P
HLR1 motif
NblS
RpaB
Strong Repression
9060300O.D.≒0.1
LB 30 mLLB 3~5 mL
Dark, 1.5 hDark, overnight Light (23~29 μmol m-2 s-1) (min)
Addition LB 1 mL
Dark, 1 h
RpaB"æ³)ï&��ßá!�.hÆÁë�Ö"¶Ô2è'��RpaB!��.>X:Y6EY?#NblS��.���Ï�/��.�(�NblSx$RpaB2��Ç!ÆÁ��.��)!�RpaB!êí�/.hliAJWPY@Y"\¼!ÉÇöb�ā°ÍÐ�#GFPĂ�ôÚ�/.+� JS=MD2dl�� (A) �Zª�NblS#�h!c��Ä�.øv=CV=*h|¡VDA8="�v!+-ìÓ�/.zÝ�)¤§�/��.�(�hÃ�2Ê¥êí�.o">X:Y6EY?Cph1!)ËÉ��N¯Ñ"hêíDO4X#Cph1�C¯Ñ"6EY?DO4X#NblS���6OS>X:Y2ÆÁ��.JS=MDāBĂ)dl���ßá!�i���¬→«!+.h±aāCĂ��ÿ����0�yª"Ö�h½�!+.ÉÇöb�"ÆÁë��Z�"VKU�µl�/ (D) �RpaB"ÆÁ�TXøv¿ "�v)Îê� � (E) �����¬±a�"£q�]wm����-�ë�À)Ā� ������,�h½�±a*ë�"@4MX9�JWPY@Y÷n"¨� ��±a"�, .õv��ä��.�Ü,/��
25
+Phos-tag
-Phos-tag
CBB
0 1 5 603015
(kDa)
RpaB-P RpaB
50
37
25
20
(min)
310 483
hliA
nblA
rpoD3
rnpB
rRNA
310 483
0 1 5 603015(nt)
1,049 1,517
483 575
(min)
RpaB
25
+Phos-tag
-Phos-tag
0 1 5 603015
(kDa)
RpaB-P
CBB
50
37
25
20
(min)
hliA
nblA
rpoD3
rnpB
rRNA
483 575
1,049 1,517
310 483
0 1 5 603015
310 483
(nt) (min)
310 483
1,049 1,517
hliA
nblA
rpoD3
rnpB
rRNA
310 483
0 1 5 603015(nt)
483 575
(min)
RpaB
25
+Phos-tag
-Phos-tag
0 1 5 603015
(kDa)
RpaB-P
CBB
50
20
25
37
(min)
Dark Low Light
Dark Light Dark Light
Dark Low Light
Light Strong Light Light Strong Light
(B)
(C)
(D)
(A)
(B)
(C)
(Acknowledgments) This study was supported by Daichi INAMI, Akira YASUDA and Tomoko NISHIMURA (Chiba Univ.).
9060300O.D.≒0.1
LB 30 mLLB 3~5 mL
Dark, 1.5 hDark, overnight Light (23~29 μmol m-2 s-1)
����D→L���� �� �� 200528 ���
(min)
Addition LB 1 mL
Dark, 1 h
GFP(804 bp)
9060300
CphI-NblS
rRNA
9060300 9060300(min)
CphI-ShortNblS-Full
1,049
575
1,517(nt)
����D→L���� ������ �200603 ����
RpaB(+Phos-tag)
9060300
CBB stain
NblS-Full
RpaB(-Phos-tag)
7942CL 9060300
CphI-NblS9060300
CphI-Short
25
(kDa)
RpaBRpaB-P
75
50
37
25
(min)
(D) (E)
(Yasuda et al., 2020)