Restriction and modification of bacteriophages by R+ strains of Escherichia coli K12

Restriction and modification of bacteriophages by R+ strains of Escherichia coli K12

Vol. 30, No. 6. 1968 BIOCHEMICAL RESTRICTION AND BIOPHYSICAL RESEARCH COMMUNICATIONS AND MODIFICATION BY R+ STRAINS D. OF OF BACTERIOPHAGES ...

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Vol. 30, No. 6. 1968

BIOCHEMICAL

RESTRICTION

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

AND MODIFICATION

BY R+

STRAINS

D.

OF

OF BACTERIOPHAGES

ESCHERICHIA

Bannister

and

S.

COLI

W.

K12

Glover

M.R.C. Microbial Genetics Research Hammersmith Hospital, Ducane Road,

Received

February It

Watanabe (R

al.

factors) that

l5

K12

strain

for

of

the

W31,

T3 The

of

suitable

$1.

the

plating

151

or

more

the

basis that

of of some

of

Groups BF23

and

two

groups

resistance

had

e.o.p. these

VI

a reduction been

(Meynell,

These values groups

and in

Ix e.o.p.

previously personal

show

~2,

PI,

Dr.

Naomi

and

6 others)

can

be

-fi'

and

phage

W31.

by

results

both of

cultures.

the

I.

e.o.p.

R+

accurately

Table

communication).

h, $80,

coli

hundred

the

less

-fi+ than

Six

colicin

of

six

efficout

this

survey.

restricted into

ten

interesting

groups to

R factors. 1 x

10B3

the

R factors

production All

plating

divided is

fold

The

of

It

- fi-

Escherichia

serial

in

for

the

spotting

of

the

65 fi-

by

shown

tested

to

I

(7lfi",

phages:

R factors

for

confined

R factors

59

an

co1

by

20 fi'

contain

with

provided

lawns

fi',

(1966)

kindly

1 presents

(33

not

in

the

measured

Table 59

151

factors

were out

then

transfer

associated

determined)

strains

(19641,

Siccardi

was

of

of

onto

Okada

resistance

was

this

not

carried

phage.

phages.

the

that

W31

a total

were

tested,

the

and

phage

(e.o.p.1

R factors

and

restriction

was

of

and

bacteriophages.

BF23

J5-3R*

Watanabe

class

certain

was

for

grown

of

fi-

surveyed

The

55-3

of

phages

survey

dilutions

the

-met

by

the

character

pro

and

that

R factors,

fi

initial

of

iencies

F-

reported

growth

by

which

dilutions

note

the

We have

55-3

Datta.

on

1966)

alone.

and

one

(1964,

conferred

R factors

Of

previously

restriction

resistance

BF23,

been

restrict

showed

W.12.

23, 1968

has

--et

Unit, LONDON,

and were

for

phage in

colicin

colicinogenic

these

X

The e.o.p. morphology,

others

Ix

The is

-I

1.0

1.0

4 io-3

lx10

5x1o-1

1.0

1.0

1.0

10.

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

1.0

6x10-l

Naomi

Datta

experiments cases where,

1.0

1.0

1.0

1.0

1.0

< 10' 2

* 10-3

1.0

3x10-l

1.0

3x1o-2

7x10-3

< 10' 2

2

2

4 lo-

-t lo-

2x10-

of

E.

1.0

1.0

1.0

1.0

and

Dr.

55-3

Elinor

Meynell.

a group. plaque

51

32

3

1

2

--

3

6

1

9

2

2

1

-

1

-

-

-

of R factors fi+ not tested

4

*No.

17

6

2

-

1

3

1

3

-

fi-

member of of abnormal

7x10-7

1.0

1.0

1.0

1.0

7x1o-4

lxlo-l

Pl

coli

with each because

3x10-5

2

1x10- 4

4xlo-4

strains

P2

R+

$80

on

Xvir

bacteriophages

1.0

1.0

1.0

1.0

BF23

of

1.

e.o.p. values are based on averages of at least two indicated as less than ( 4) a certain value in those plaque counts were subject to considerable error. *The - fi character of the R factors was determined by Dr.

1.0

1.0

1x1o-1

1x10' 1

1.0

8x10-l

5x10-l

10-3

7x10-l

<

7x10-l

< 10-3

lx1o-1

plating

-=i 10-3

-=i 10-3

1.0

1.0

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1.0

of

10-3

<

7x10-3

8x10'~

VII

VIII

< 10-3

< 10 -3

VI

1.0

1.0

1.0

1.0

w31

< 10-3

<

1.0

1.0

1.0

1.0

T3

10-3

V

IV

III

II

I

R factor group

Efficiency

TABLE

Vol. 30, No. 6. 1968

for

-co1

co1

I

Ib

and

BIOCHEMICAL

-co1

resistance

I

in

Table

resistant.

has

A number 1,

also

of less

8 x IO- 5.

More

commonly

host

is

(1966)

Arber

1

3 x lothere

, and

is

(T3,

phages

after

host

of

e.0.p.

and

reduction

in

than

Tl

1 x

isolate

are

size,

in

is no

halo). the

due

to

--et

cited

the

This

effect

phage

was

host

is

more

likely

one

or

more

of

(1966)

restricting However,

an e.o.p. in

expected and

if,

subsequent

reduction

factors,

in

for

slightly

already

specific (e.g.

be

the

several

1

female

not

that

of

Table

modified

and

have

on

to

morphology

would

It

IO-

so-called

plaque

adsorption,

al.

with

reported

1

1957).

examples

in

BFZj

x

phage

with

with

a variation

and

on B (PI) the

and,

associated

(I

(Lederberg

In

size

$1

modified

plates

inefficient

Watanabe

for

plaque

W31, high

host

it.

normal.

size

burst

by

T3,

units

fd.B

infection,

were

transmission. size

of

infection plaque

phage

or

round

of

phages

(1966).

exceptionally

log.

there

halo,

are

these

Siccardi phages

e-0-p.

modified

$I),

by

several

in

the

first

rounds

that

of

for

1.0, the

reduction

of the

than

shown

W31 and

crenelation

found values

by

is

some

been

reduced

has

Restriction

e.o.p.

though

strains

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

example

impaired

reported

a reduced

burst

an R+ host.

In

many

lo-'

and

mutants,

instances

where

the

phage

is

which

are

no

the not

e.o.p.

host

recorded

modified

longer

restricted,

host

modification

in

it

the

has

from

Table

been

1 is

possible

plaques

less to

with

normal

morphology. We have group is

I, group

and

of

demonstrated h,

specific

in

group

II

(and

vice

group

II

now

plates

In

addition,

plate II

$80

the

which reported

be

identical

host

modify by

them

that

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in

phage

versa)

and

phages

h,

on the with x,

and

may belong

group

and

other the an

on in

on $80

Pl

Host group

that

any

other

on

one

which of

is

our

737

by

$80

of

Table

--et does

in phages

of

member

I

VI.

these

any

group

but

P2

by

group,

Watanabe

and

restricted

the

(see

V or

of

on

either

restricts groups

A,

still

grown

groups found

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I

member

grown

phages

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phage

restricting

R factors

to

II.

grown also

efficiently

inefficiently may

and

of

(Table

or

II,

still Group

I).

a1.(1964, not

2).

modify

1966) x

Vol. 30, No. 6. 1968

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

Table Host

modification

of

phages

h,

2.

$80,

PI

and

P2

in

E.

coli

J5-3R+

strains

R factor

Phage

none

group (1

h.J5-3

2x1o-2 lX1O-2

1.0

1.0

h.

group

II

1.0

2x10'

1.0

1x10'

4

group

1

1.0

1.0

$80.

group

II

1.0

1x10-

1.0

3x10-5

1.0

1.0

PI.

55-3

PI.

group

The e.o.p. members of

for the

I

II The phages group.

1.0

4

$80.

group

(9 R factors)

4x1o-4

1

P2.

II

1.0

group

P2.J5-3

group

R factor)

h.

$80.55-3

I

7x10-3 4x10-3

4

1.0

1.0

7x1o-4

1.0

1.0

figures grown

in the on group

table represent II are expressed

approximate as an

e.o.p. average for

values. all

Acknowledgements The the generous advice and Scholarship

authors wish to thank Dr. Naomi Datta and Dr. Elinor Meynell gift of the R factors used in this work and for the unstinted unpublished information concerning them. One of us (D-B.) wishes to thank the Medical Research Council for for Training in Research Methods.

References Arber, W. (1966) J. Mol. Biol. 20, 483. Lederberg, S. (1957) Virology 3, 496. Siccardi, A. G. (1966) Genet. Res. Camb. 8, '1. and M. Okada (1964) J. Bacterial. Watanabe, H. Nishida, C. Ogata, T. Arai Watanabe, T., 88,

Watanabe,

and

87, S.

727. Sato

(1964)

J.

Bacterial.

716. T.,

!z,

219.

477.

T.

Takano,

T.

Arai,

H.

Nishida

and

S.

Sato

(1966 ) J.

Bacterial.

for

a