\ List of the Projects of Experiments.

(Oct. 4, 1955)
Tetsuo Tino

I. Experiments on the effect of the difference of phase on the H-antigen transduction.

1. Reconstruction experiment.

b:(1,2) Gal™ (1):enx Galt 4:1,2 Gal
24hour Penasgey broth 24hour penageay broth 24hour penagsay broth
culture. /m2 culture. 109/ml culture. 107/ml

dilute ~—~cheek conc. — dilute check conc.
of cultures of caltures.
& x 107/m1 4 x'10?/m1

\

5 ml 0.25 ml 0.5 1
aout 33 jai 5x 108/m1)

A

 

 

a
brush on anti-i,-1,2 MGA
check number of the cells in a loop plates 31) large loop
(8 x 107/ml)
“3: (1,2) Gal” (i):enx Gal i:1,2 Gal’
8/ brush 8/ brush 4 x 106/brush

2. Transduction frem Sa}. abony(Fla, bsenx) to TM-2 Fla--5 (1:12).
a). Obtain Gal~ mtant from TM-2 Fia*-s.
b). Obtain single phase culture of TM-2 Fla -5.

eo}. Traneduction experiment:
TH-2 Fla™-5, 43(1,2)

Sal. abony Plat, bv: (enx) ———_—-g 1:1 mixture of Gal.
T™N-2 Fla P25, (4)1,2

Gal

Sal. bony Fla’? (b) $enx x f ft

Screen Fla’ swarms on MGA-plates, ani test antigen.
3. Transduction between single phase cultures (antiserum selection).

a). [= 4 ——~£ 1:1 mix of Sal. abony b, Gal” and enx, Gal’.

 

TM-2 1,2 nS " " " °
b). {M2 4 x , b, Gal’ and enz,Gal .
M-2 1,2——x " " " .

x l:l mix of ™-2 1, Gal and 1,2, Gal*.

 

c). Sal. abony b

" enx ——-—-x 8 " 8 .

x " 4, Gal* and 1,2,2Gal™.

 

a). JSal. abony b

" enx —————"-x " " Cl °

II. Experimants on the i:b: ens-etkehere
4, Recover unstable i°b*/ a*o” from original culture of i:b;enx-strain.

5. Compare changed reactivity of b and i in parent cultures as function of mediun.

™-2 1

Sal. aboxy > Penassay broth
" " p,i transfer to EMB-ager
" "  (b) 4 Minimal agar
u iy L

Compare the intensities of agglutination for anti-b and anti-i.

6. Alternative phase of ity,
a).Brush the culture of i*b” on MGA containingganti-i or anti~i + anti-enx,

b). Test the antigen types of the swarns developed.

%. Transductton of H, from 1:b:enx.
a}. Prepare lysate of i:b:enx. If U.¥. is not efficient to induce phage,

use penicillin.

 

b). Transduction: i:r:enx x Sal. abony 3:enx (select by anti-b and ~ept)

 

c). Transduction: Plati:r:enx x Sal. paratyphi SW666 Fla »
8. Test of the locus specificity in H,-transduction to i:r:enz.

x itrsenx

 

a). Sal. sendai a:1,5
b). Sal. altendorf o:1,7 -——x "
c). Seal. sega a:36 Hx *

In each case, brush on MGA containing anti~-r and anti-enx, and test the types

of antigen of swarms developed,