*��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

!����

���#��

� ������

�"���Promoter

����

(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)