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제한효소의 Star 활성

제한효소의 star activity

특정 반응 조건에서 제한효소는 기질 DNA에 대한 특이성을 잃고 원래 인식부위와는 다른 염기서열을 절단하는 경우가 있다. 이러한 현상을 ‘star activity’ 라고 부르며, 효소, 기질 DNA, 반응 조건에 따라 대부분의 제한효소가 이러한 star activity를 가질 수 있다고 알려져 있다. 또한 인식 부위 이외의 서열을 절단하는 것 이외에도 ‘nicking activity’ (부분적으로 절단된 DNA)가 나타날 수도 있다.
Star activity를 억제하기 위해서는 글리세롤 농도를 낮추고, 중성 pH를 유지하며 염 농도를 높이는 조건에서 반응을 수행하는 것이 좋다. 하지만 이러한 조건으로 인해 반응성이 낮아질 수도 있다. 이러한 특정 실험 조건에서 star activity가 나타날 수 있는 제한효소와 그 경우 인식하는 서열은 아래 표에서 확인할 수 있다 (인식서열 정보 및 반응 조건은 표 하단 참조).

Restriction enzyme	Normal recognition sequence	Reaction conditions*	Recognition sequence**	Reference
Aat II	GACGT↓C	E		Takara Bio Inc., unpublished observations
Aor13H I	T↓CCGGA	A, C, E		Takara Bio Inc., unpublished observations
BamH I	G↓GATCC	A, B, E, F	GRATCC GGNTCC GGANCC GGATYC	Nath and Azzolina 1981; George, Blakesley, and Chirikjian 1980; George and Chirikjian 1982; Malyguine, Vannier, and Yot 1980
Ban II	GRGCY↓C	A, E, F		Takara Bio Inc., unpublished observations
BciT130 I	CC↓WGG	A, E,
Bgl I	GCCNNNN↓NGGC	F		Takara Bio Inc., unpublished observations
Bgl II	A↓GATCT	E		Takara Bio Inc., unpublished observations
Bln I	C↓CTAGG	B, E, G
BmeT110 I	C↓YCGRG	E
BspT104 I	TT↓CGAA	A, E		Takara Bio Inc., unpublished observations
BstP I	G↓GTNACC	A, F		Takara Bio Inc., unpublished observations
Bst1107 I	GTA↓TAC	F		Takara Bio Inc., unpublished observations
Dde I	C↓TNAG	A, B, E
EcoO65 I	G↓GTNACC	E, F		Takara Bio Inc., unpublished observations
EcoR I	G↓AATTC	A, B, E, F	NAATTN	Malyguine, Vannier, and Yot 1980; Polisky et al. 1975; Tikchonenko et al. 1978; Hsu and Berg 1978; Woodbury, Hagenbuchle, and von Hippel 1980
EcoR V	GAT↓ATC	E	RATATC GNTATC GANATC GATNTC GATANC GATATY	Halford, Lovelady, and McCallum 1986; Takara Bio Inc., unpublished observations
EcoT22 I	ATGCA↓T	F, H		Takara Bio Inc., unpublished observations
Fba I	T↓GATCA	A, C, E, F		Takara Bio Inc., unpublished observations
Hae III	GG↓CC	A		Nath and Azzolina 1981
Hha I	GCG↓C	A, E		Malyguine, Vannier, and Yot 1980
Hinc II	GTY↓RAC	E		Takara Bio Inc., unpublished observations
Hind III	A↓AGCTT	B, E	RAGCTT ANGCTT AAKCTT AAGMTT AAGCNT AAGCTY	Hsu and Berg 1978; Nasri and Thomas 1986; Takara Bio Inc., unpublished observations
Hpa I	GTT↓AAC	A, E		Nath and Azzolina 1981; Takara Bio Inc., unpublished observations
Kpn I	GGTAC↓C	A, B, C, E		Takara Bio Inc., unpublished observations
Mun I	C↓AATTG	A, F		Takara Bio Inc., unpublished observations
Nco I	C↓CATGG	A, E		Takara Bio Inc., unpublished observations
Nde I	CA↓TATG	A, B, G
Nhe I	G↓CTAGC	A, C, E, F		Takara Bio Inc., unpublished observations
Nsb I	TGC↓GCA	B		Takara Bio Inc., unpublished observations
PshB I	AT↓TAAT	D		Takara Bio Inc., unpublished observations
Psp1406 I	AA↓CGTT	E		Takara Bio Inc., unpublished observations
Pst I	CTGCA↓G	A, E		Nath and Azzolina 1981; Malyguine, Vannier, and Yot 1980; Takara Bio Inc., unpublished observations
Pvu II	CAG↓CTG	A, E	NAGCTG CNGCTG CANCTG CAGNTG CAGCNG CAGCTN	Nasri, Sayadi, and Thomas 1985; Nasri and Thomas 1987; Takara Bio Inc., unpublished observations
RspRS II	T↓TAA	A
Sac I	GAGCT↓C	A, E		Takara Bio Inc., unpublished observations
Sal I	G↓TCGAC	A, E		Malyguine, Vannier, and Yot 1980; Takara Bio Inc., unpublished observations
Sau3A I	↓GATC	A, E	SATC GMTC GAKC GATS	Pech, Streeck, and Zachau 1979; Takara Bio Inc., unpublished observations
Sca I	AGT↓ACT	B, C, F		Grosskopt, R. and Kessler, C., unpublished observations
Sfi I	GGCCNNNN↓NGGCC	B, E		Takara Bio Inc., unpublished observations
Smi I	ATTT↓AAAT	A, B, C, E		Takara Bio Inc., unpublished observations
Spe I	A↓CTAGT	E, F		Takara Bio Inc., unpublished observations
Sse8387 I	CCTGCA↓GG	E, F		Takara Bio Inc., unpublished observations
Ssp I	AAT↓ATT	A, C, E, F		Takara Bio Inc., unpublished observations
Taq I	T↓CGA	A, B, E, G		Takara Bio Inc., unpublished observations
Tth111 I	GACN↓NNGTC	B, C	NACNNNGTC GNCNNNGTC GANNNNGTC GACNNNNTC GACNNNGNC GACNNNGTN	Shinomiya et al. 1982
Van91 I	CCANNNN↓NTGG	A		Takara Bio Inc., unpublished observations
VpaK11B I	G↓GWCC	A, C, E, F		Takara Bio Inc., unpublished observations
Xba I	T↓CTAGA	A, E		Nath and Azzolina 1981; Malyguine, Vannier, and Yot 1980

*Reaction conditions:
A: high levels of glycerol
B: in the presence of Mn²⁺
C: alkaline pH
D: low pH
E: in the presence of DMSO
F: low ionic strength
G: high ionic strength
H: in the presence of 2-mercaptoethanol
**Recognition sequences:
M: A or C
K: G or T
N: A or C or G or T
R: A or G
Y: C or T
W: A or T
S: G or C

References
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