Preparation of ribsomal subunits by gel filtration
Date
2016-07-20
Authors
Bhoolia, Deena
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
An attempt was made to separate ribosomal subunits by gel filtl'ation
on Trisacryl GF2000 and Sepharose 4B. Trisacryl GP~2000,a sYllthet'ic
gol, , separated rat .ljver rlbosonal subunits orl 'f~'135,em column with a
resolution of ""Or3, resulting 'in <'1" 60 rJ impm''ity 'of each of the
\~
subunits.
subunits" were not resolved. Sepharose 4131 an agarose based gol,
separated " the subunits by adsorpttcn chromatography rathr.r than 'i>.V
() Q
':) At 4°C, the 405 .subunf ts were eluted with a kd
()
ruO,:~9( but the GOrf' $ubud"its adsorbed to th~.'g,~Jl and were eluted I;
/)
when the temperature of "the column was increas~d to 250C..3SoC.
(' ,. ('-. This edsorpt ion phenonenon seems to b{~ tl propert~ of a 11 agllrose
-: \') ~\:;
based gels studied here, includ'ing Sepharose 2.B and .seph'ato~eoBt arid
is exc 1us iva to !,mamm'i'lan r ibosOIntrt subuni ts • Anal,ys1S 0'(" the
, u
subunits by in vitro r14C]polypheny1alanine sy'nthes'is showed OCI
" ,,'c. /\ I 7', " diff'erence 'in the act lvtt ies of dbosoma'J ~ubunH~ p~epdrf#d by
;/'/ '
grndient centrifugation or by 5epharos,e cni"OmatogNPPY;' Analy~ii ~~of
"
(~ ((
the subunits by ~crylamidec'ugarose coU)po5it~) gs1s resulted ,in the
'/
resolution ,.:n"f subunits isolated fr'oill lower organisms in
o II
non-denaturlnq !Jcl systems and SI,lbut1its from m«mm~nan'tissue in
II
/'\';1
Ga'! f'iltrat'ion does tyffer a 5t!i:l;~'ble metrKld'¥or the pr~p;n~¥tiol1of
<::) . I \) \\ I)
ribosoma 1 'subunits I but only' if 'the act~orption JH'OrJ&~"t'Jo,s of
,) (,) ':'
o ,1)
,{
Description
A Oiss{!y'tation $uhm'itted in fulfilment
of the r~;~airements, for' the degree of
f·1aster· oi~'j Science at the University
of the ~titwatersrand '
Juhanoasbul"g
January 1991