[Biojava-l] RestrictionEnzymeManager can't correctly handle incomplete enzymes

Wed Jun 22 06:09:01 EDT 2005

(I'm not an expert on restriction enzymes.)

I was talking about AacI, of which BamHI is an isoschizomer. The recognition
site of AacI is unknown, but the one from BamHI is known. Maybe
RestrictionEnzymeManager uses the cutlocation of BamHI when asking the
unknown cutlocation of AacI.

http://rebase.neb.com/rebase/enz/AacI.html

That might also be the reason why RestrictionEnzymeManager requires links
between restriction enzymes. If a restriction enzyme entry is removed from
the REBASE file RestrictionEnzymeManager fails to read in some cases.

But I think using cutlocation of isoschizomers is wrong. Because of this:

REBASE says: "A isoschizomers is a restriction enzymes that recognize the
same DNA sequence. The cut sites may or may not be identical."

So the cut site might be different between different isoschizomers.

I searched for examples in the REBASE file, and found them:

<1>BspKT6I

<2>MboI,AspMDI,AsuMBI,Bce243I,Bfi57I,BfiSHI,BfuCI,Bme12I,Bme2494I,BsaPI,BscF
I,BsmXII,BspI,Bsp9I,Bsp18I,Bsp49I,Bsp51I,Bsp52I,Bsp54I,Bsp57I,Bsp58I,Bsp59I,
Bsp60I,Bsp61I,Bsp64I,Bsp65I,Bsp66I,Bsp67I,Bsp72I,Bsp74I,Bsp76I,Bsp91I,Bsp105
I,Bsp122I,Bsp135I,Bsp136I,Bsp138I,Bsp143I,Bsp147I,Bsp2095I,BspAI,BspFI,BspJI
,BspJ64I,BsrMI,BsrPII,BssGII,Bst19II,Bst1274I,BstEIII,BstENII,BstKTI,BstMBI,
BstXII,BtcI,Bth84I,Bth211I,Bth213I,Bth221I,Bth945I,Bth1140I,Bth1141I,Bth1786
I,Bth1997I,BthCanI,BtkII,Btu33I,Btu34I,Btu36I,Btu37I,Btu39I,Btu41I,CacI,CcoP
31I,CcoP76I,CcoP84I,CcoP95II,CcoP219I,CcyI,CdiCD6II,ChaI,Cin1467I,CjeP338I,C
paI,CpfI,CpfAI,Csp5I,Cte1179I,Cte1180I,CtyI,CviAI,CviHI,DpnII,EsaLHCI,FnuAII
,FnuCI,FnuEI,Gst1588II,HacI,HpyAIII,HpyHPK5II,Kzo9I,LlaAI,LlaDCHI,LlaKR2I,Ls
p1109II,Mel3JI,Mel5JI,Mel7JI,Mel4OI,Mel5OI,Mel2TI,Mel5TI,MeuI,MgoI,MjaIII,Mk
rAI,MmeII,Mmu5I,MmuP2I,MnoIII,MosI,Msp67II,MspBI,MthI,Mth1047I,MthAI,NciAI,N
deII,NflI,NflAII,NflBI,NlaII,NlaDI,NmeCI,NphI,NsiAI,NspAI,NsuI,Pei9403I,PfaI
,Pph288I,RalF40I,Rlu1I,SalAI,SalHI,Sau15I,Sau6782I,Sau3AI,SauCI,SauDI,SauEI,
SauFI,SauGI,SauMI,SinMI,SmiMBI,SsiAI,SsiBI,Ssu211I,Ssu212I,Ssu220I,R1.Ssu247
9I,R2.Ssu2479I,R1.Ssu4109I,R2.Ssu4109I,R1.Ssu4961I,R2.Ssu4961I,R1.Ssu8074I,R
2.Ssu8074I,R1.Ssu11318I,R2.Ssu11318I,R1.SsuDAT1I,R2.SsuDAT1I,SsuRBI,Sth368I,
TrsKTI,TrsSI,TrsTI,TruII,Tsp133I,Uba4I,Uba59I,Uba1101I,Uba1177I,Uba1182I,Uba
1183I,Uba1204I,Uba1259I,Uba1317I,Uba1323I,Uba1366I,Vha44I

<3>GAT^C

<4>2(6)

<5>Bacillus species KT6

<6>N.I. Matvienko

<7>

<8>Shapovalova, N.I., Zheleznaja, L.A., Matvienko, N.I., (1993) Nucleic
Acids Res., vol. 21, pp. 5794.

Shapovalova, N.I., Zheleznaya, L.A., Matvienko, N.I., (1994) Biokhimiia,
vol. 59, pp. 1730-1738.

<1>MboI

<2>AspMDI,AsuMBI,Bce243I,Bfi57I,BfiSHI,BfuCI,Bme12I,Bme2494I,BsaPI,BscFI,Bsm
XII,BspI,Bsp9I,Bsp18I,Bsp49I,Bsp51I,Bsp52I,Bsp54I,Bsp57I,Bsp58I,Bsp59I,Bsp60
I,Bsp61I,Bsp64I,Bsp65I,Bsp66I,Bsp67I,Bsp72I,Bsp74I,Bsp76I,Bsp91I,Bsp105I,Bsp
122I,Bsp135I,Bsp136I,Bsp138I,Bsp143I,Bsp147I,Bsp2095I,BspAI,BspFI,BspJI,BspJ
64I,BspKT6I,BsrMI,BsrPII,BssGII,Bst19II,Bst1274I,BstEIII,BstENII,BstKTI,BstM
BI,BstXII,BtcI,Bth84I,Bth211I,Bth213I,Bth221I,Bth945I,Bth1140I,Bth1141I,Bth1
786I,Bth1997I,BthCanI,BtkII,Btu33I,Btu34I,Btu36I,Btu37I,Btu39I,Btu41I,CacI,C
coP31I,CcoP76I,CcoP84I,CcoP95II,CcoP219I,CcyI,CdiCD6II,ChaI,Cin1467I,CjeP338
I,CpaI,CpfI,CpfAI,Csp5I,Cte1179I,Cte1180I,CtyI,CviAI,CviHI,DpnII,EsaLHCI,Fnu
AII,FnuCI,FnuEI,Gst1588II,HacI,HpyAIII,HpyHPK5II,Kzo9I,LlaAI,LlaDCHI,LlaKR2I
,Lsp1109II,Mel3JI,Mel5JI,Mel7JI,Mel4OI,Mel5OI,Mel2TI,Mel5TI,MeuI,MgoI,MjaIII
,MkrAI,MmeII,Mmu5I,MmuP2I,MnoIII,MosI,Msp67II,MspBI,MthI,Mth1047I,MthAI,NciA
I,NdeII,NflI,NflAII,NflBI,NlaII,NlaDI,NmeCI,NphI,NsiAI,NspAI,NsuI,Pei9403I,P
faI,Pph288I,RalF40I,Rlu1I,SalAI,SalHI,Sau15I,Sau6782I,Sau3AI,SauCI,SauDI,Sau
EI,SauFI,SauGI,SauMI,SinMI,SmiMBI,SsiAI,SsiBI,Ssu211I,Ssu212I,Ssu220I,R1.Ssu
2479I,R2.Ssu2479I,R1.Ssu4109I,R2.Ssu4109I,R1.Ssu4961I,R2.Ssu4961I,R1.Ssu8074
I,R2.Ssu8074I,R1.Ssu11318I,R2.Ssu11318I,R1.SsuDAT1I,R2.SsuDAT1I,SsuRBI,Sth36
8I,TrsKTI,TrsSI,TrsTI,TruII,Tsp133I,Uba4I,Uba59I,Uba1101I,Uba1177I,Uba1182I,
Uba1183I,Uba1204I,Uba1259I,Uba1317I,Uba1323I,Uba1366I,Vha44I

<3>^GATC

<4>2(6)

<5>Moraxella bovis

<6>ATCC 10900

<7>ACFGKNQRUVX

<8>Anton, B.P., Brooks, J.E., Unpublished observations.

Gelinas, R.E., Myers, P.A., Roberts, R.J., (1977) J. Mol. Biol., vol. 114,
pp. 169-179.

Huang, L.-H., Farnet, C.M., Ehrlich, K.C., Ehrlich, M., (1982) Nucleic Acids
Res., vol. 10, pp. 1579-1591.

Ueno, T., Ito, H., Kimizuka, F., Kotani, H., Nakajima, K., (1993) Nucleic
Acids Res., vol. 21, pp. 2309-2313.

Ueno, T., Ito, H., Kotani, H., Nakajima, K., Japanese Patent Office, 1993.

<1>Mel3JI

<2>MboI,AspMDI,AsuMBI,Bce243I,Bfi57I,BfiSHI,BfuCI,Bme12I,Bme2494I,BsaPI,BscF
I,BsmXII,BspI,Bsp9I,Bsp18I,Bsp49I,Bsp51I,Bsp52I,Bsp54I,Bsp57I,Bsp58I,Bsp59I,
Bsp60I,Bsp61I,Bsp64I,Bsp65I,Bsp66I,Bsp67I,Bsp72I,Bsp74I,Bsp76I,Bsp91I,Bsp105
I,Bsp122I,Bsp135I,Bsp136I,Bsp138I,Bsp143I,Bsp147I,Bsp2095I,BspAI,BspFI,BspJI
,BspJ64I,BspKT6I,BsrMI,BsrPII,BssGII,Bst19II,Bst1274I,BstEIII,BstENII,BstKTI
,BstMBI,BstXII,BtcI,Bth84I,Bth211I,Bth213I,Bth221I,Bth945I,Bth1140I,Bth1141I
,Bth1786I,Bth1997I,BthCanI,BtkII,Btu33I,Btu34I,Btu36I,Btu37I,Btu39I,Btu41I,C
acI,CcoP31I,CcoP76I,CcoP84I,CcoP95II,CcoP219I,CcyI,CdiCD6II,ChaI,Cin1467I,Cj
eP338I,CpaI,CpfI,CpfAI,Csp5I,Cte1179I,Cte1180I,CtyI,CviAI,CviHI,DpnII,EsaLHC
I,FnuAII,FnuCI,FnuEI,Gst1588II,HacI,HpyAIII,HpyHPK5II,Kzo9I,LlaAI,LlaDCHI,Ll
aKR2I,Lsp1109II,Mel5JI,Mel7JI,Mel4OI,Mel5OI,Mel2TI,Mel5TI,MeuI,MgoI,MjaIII,M
krAI,MmeII,Mmu5I,MmuP2I,MnoIII,MosI,Msp67II,MspBI,MthI,Mth1047I,MthAI,NciAI,
NdeII,NflI,NflAII,NflBI,NlaII,NlaDI,NmeCI,NphI,NsiAI,NspAI,NsuI,Pei9403I,Pfa
I,Pph288I,RalF40I,Rlu1I,SalAI,SalHI,Sau15I,Sau6782I,Sau3AI,SauCI,SauDI,SauEI
,SauFI,SauGI,SauMI,SinMI,SmiMBI,SsiAI,SsiBI,Ssu211I,Ssu212I,Ssu220I,R1.Ssu24
79I,R2.Ssu2479I,R1.Ssu4109I,R2.Ssu4109I,R1.Ssu4961I,R2.Ssu4961I,R1.Ssu8074I,
R2.Ssu8074I,R1.Ssu11318I,R2.Ssu11318I,R1.SsuDAT1I,R2.SsuDAT1I,SsuRBI,Sth368I
,TrsKTI,TrsSI,TrsTI,TruII,Tsp133I,Uba4I,Uba59I,Uba1101I,Uba1177I,Uba1182I,Ub
a1183I,Uba1204I,Uba1259I,Uba1317I,Uba1323I,Uba1366I,Vha44I

<3>GATC

<4>

<5>Megasphaera elsedenii 3J

<6>P. Pristas

<7>

<8>Piknova, M., Filova, M., Javorsky, P., Pristas, P., (2004) FEMS
Microbiol. Lett., vol. 236, pp. 91-95.

Piknova, M., Pristas, P., Javorsky, P., (2004) Folia Microbiol. (Praha),
vol. 49, pp. 191-193.

-----Oorspronkelijk bericht-----

Van: mark.schreiber at novartis.com [ <mailto:mark.schreiber at novartis.com>
mailto:mark.schreiber at novartis.com]

Verzonden: woensdag 22 juni 2005 11:25

Aan: Jesse

CC: biojava-l at biojava.org; biojava-l-bounces at portal.open-bio.org

Onderwerp: Re: [Biojava-l] RestrictionEnzymeManager can't correctly handle
incomplete enzymes

I take your point but I notice that BamHI is an isoscizomer. Is the cleavage
site of BamHI really unknown??

- Mark

"Jesse" <jesse-t at chello.nl>

Sent by: biojava-l-bounces at portal.open-bio.org

06/22/2005 04:15 PM

        To:     <biojava-l at biojava.org>

        cc:     (bcc: Mark Schreiber/GP/Novartis)

        Subject:        [Biojava-l] RestrictionEnzymeManager can't correctly
handle     incomplete 

enzymes

RestrictionEnzymeManager can't correctly handle incomplete enzymes and gives
wrong data.

(Correct me if I'm wrong.)

I'm not sure if this is already discussed or not.

I think RestrictionEnzymeManager can not handle incomplete restriction
enzymes.

BioJava 1.4Pre2 knows two types of RestrictionEnzymes:

-RestrictionEnzyme.CUT_SIMPLE

-RestrictionEnzyme.CUT_COMPOUND

But in REBASE, there are also other restriction enzyme entries:

-Unknown recognition sites. For example "<3>?". RestrictionEnzymeManager
skips this one (which is ok).

-Unknown cut location. For example AacI "<3>GGATCC".

The problem with RestrictionEnzymeManager is with those REBASE entries which
have an unknown cutlocation. RestrictionEnzymeManager  will actually tell
that there is a cutlocation, even though it's unknown in the REBASE file.

For example:

http://rebase.neb.com/rebase/link_withrefm

--------- REBASE ENTRY -----------

<1>AacI

<2>BamHI,AaeI,AcaII,AccEBI,AinII,AliI,Ali12257I,Ali12258I,ApaCI,AsiI,AspTII,

Atu1II,BamFI,BamKI,BamNI,Bca1259I,Bce751I,Bco10278I,BnaI,BsaDI,Bsp30I,Bsp46I

,Bsp90II,Bsp98I,Bsp130I,Bsp131I,Bsp144I,Bsp4009I,BspAAIII,BstI,Bst1126I,Bst2

464I,Bst2902I,BstQI,Bsu90I,Bsu8565I,Bsu8646I,BsuB519I,BsuB763I,CelI,DdsI,Gdo

I,GinI,GoxI,GseIII,GstI,MleI,Mlu23I,NasBI,Nsp29132II,NspSAIV,OkrAI,Pac1110I,

Pae177I,Pfl8I,Psp56I,RhsI,Rlu4I,RspLKII,SolI,SpvI,SurI,Uba19I,Uba31I,Uba38I,

Uba51I,Uba88I,Uba1098I,Uba1163I,Uba1167I,Uba1172I,Uba1173I,Uba1205I,Uba1224I

,Uba1242I,Uba1250I,Uba1258I,Uba1297I,Uba1302I,Uba1324I,Uba1325I,Uba1334I,Uba

1339I,Uba1346I,Uba1383I,Uba1398I,Uba1402I,Uba1414I,Uba4009I

<3>GGATCC

<4>

<5>Acetobacter aceti sub. liquefaciens

<6>IFO 12388

<7>

<8>Seurinck, J., van Montagu, M., Unpublished observations.

----------------------------------

--------- RestrictionEnzyme values --------

Name: AacI

RecognitionSite:ggatcc

ForwardRegex: g{2}atc{2}

ReverseRegex: g{2}atc{2}

CutType: 0 (RestrictionEnzyme.CUT_SIMPLE)

DownStreamEndType: 2

IsPalindromic: true

DownstreamCut: 1, 1,

-------------------------------------------

As you can see, AaCI is used as RestrictionEnzyme.CUT_SIMPLE and it has a
cutlocation while the REBASE entry says that the cutlocation is unknown,
only the recognition site is known. So RestrictionEnzymeManager should also
filter out those with an unknown cutlocation, otherwise it gives wrong data.

- Jesse

[Biojava-l] RestrictionEnzymeManager REBASE reader bug?

mark.schreiber at novartis.com mark.schreiber at novartis.com Tue Jun 21
22:22:52 EDT 2005 

Hello -

This is now checked in. All tests pass (no surprise as checking for null
never hurt anyone). This will make it into biojava1.4. If you want to add a
test to the Junit to ensure this stays fixed it would be most appreciated.

I also remember some discussion a while back about the behaivour of certain
enzymes with respect to their cleavage points which may or may not 

have been a bug. Was this ever resolved? If so does anything need fixing?

Thanks.

- Mark

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