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This Week’s Citation Classic
CCINUI~EB42
!~OB~ 21,1991
Aviv H & Leder P. Purification of biologically active globin messenger RNA by
chromatography on oligothymidylic acid-cellulose. Proc. Nat~Aco4 Sci. USA
69:1408-12, 1972.
[Laboratory of Molecular Genetics. National Ina*nute of Child Health and Human Development,
National Institutes ofHealth. Bethesda. MDI
______________________
A convenient technique for the partial punfication of
tairnng a thoit ~etth of thymkines (ollgo dl). It
large quantities of functional, poly(adenyluc acid)-rich was not obvious,, howevee, that suds a ihoit A:T
mRNA is described. Biologically active rabbit glabin hybrid would be stable. To syu1hcaLa~c ollgo dT
mRNA was punfied by this procedure and assayed for diemleally hooked to cellulose, I fowida pnscedssie
its ability to direct the syiitheais of rabbit globin in a
dut invoked aindemation using
cell-free extract of asciws tunsy. 111w Sd ® indicates used
DCC, idsidiIs veryunwitive to water. knowkigve,y
that this paper has been cited in more than 6,150
Hate orgamc d~.try, this was not encouraging
publications.l
for me. I sought advice from bead Sdiedd~of the
*5*Wesvnasn Institute who apent a _____
hi the
lab. I set up the synthesis whids itsioked a whale
How to Win Friends and Advance Science
week of shaking. I did this with great h~sitatlo~
sure that it wouldni work. This was ..v.bJlly the
Haim Aviv
5 my cases,,
only organic synthesis that I ever did in
Department of Plant Genetics
aid
it
worked.
Weivnann institute of Science
Having lots of extracted reticulocyte RNA, I set
Relsovot 76100
up the procedure that is described In this mod.citrd
Israel
paper, and it worited rigid away. We got plenty of
I came to Philip Leder’s laboratory at the National
labeled amino acids incorporated into protein ditnstitute of Child Health and Human Development
rectedbymRNA.
in 1970 after finishing my doctorate at the
I immediately followed this ~
by runWeumann ln~tituteof Science in IsraeL The main
ning through this little column of oliga d1 using
reieardl direction of the lab was to study imiTwno.
RNA preparations extracted from niyelwna cells,
globulin biosynthesis usmg molecular biology ~which, ksc*Dy, I had not discarded.This opened up
proaches. One of the first steps In this endeavor was
many studies on immunogiobin mRNA,, wisids were
to isolate the immunogiobulin mRNA from my- ——4
eloma cells (MOPC-41). The isolation ofmRNA was
Having lots of pure globin mINA., we joined
thought to be essential in order to have a handle on forces with Jeff Rose and
5 Ed Scoinids to synthesize
the biosynthesis ofimmunoglobulins.
coniplenientary DNA —ai 6.~xteta.Lmilestone hi
After establishing a cell-free translation system the development of gene doning and moleadar
usingan extract from a murme aacitescell line,’ we
set up to establish an extraction procedure foe
My procedure for mRNA purification made me
tnRNA from the myeloma cells. However~for about lots of friends. Many colleagues have asked for urnsix months we could not get any counts incorpo- pies of this precious oiigo dT celhslose, width I was
rated Irons labeled amino adds, whatever we tried. willing to sharewith then provided theymade their
It was a very frustrating experience. Then I sug- owis batch under my guidance and we ~llt It.
gested that we check out our extraction procedures
I also bad aimplaints that the column did not
(parflcularIy the quality of freshly distilled phenol)
work. It mostly turned out that the column was not
using mRNA extracted from rabbit reticulocytes for
proped~
whids there were a number of published proceIn the nearly 20 years since the procethue was
dures. This wasdone just asa control. It worked fine published, it ha been cited hi thousand,of psth&awhen mRNA was extracted from retics, but failed
dons and is still the most airiness t.,aj~ij~j~
of
when extracted from myeloma tissue.
mRNA purification. ~iy riall changes hi the proAt that time, a number of papers were published cedure subsequendy have bees hdrodeced. Some
showing that globin mRNA has a sfretd, of adenorave on thewashing step when purity Is not crucial,
2
sines at the 3’ end whose ftmction was not clear. It some recycle the unbound material Ifa higher yield
occurred to me that we might be able to separate Is desired. But to the best of my knowledge, most
giobin mRNA from ribosonsal RNA and tRNA by researdier, have been happy with the procedure as
hybridizing mRNA to a solid phase polymer con- originally described.
—
I. Aviv H, Bourn. I & Led,r P. Protein-synthesis directed by encephalomyocarditis virua-RNA—propcrues oI* transfer
RNA-drpcndenl system. Proc. Na:. Aca,L Sri. (iSA 68:2303-7. 1971 (Cited 135 times.)
2. Edmoods M & Cariniels M 6. Isolation and characterization of adenosinc monophosphate-rich polynucleoudes
synthesized by Ehrlich arctics cells. J. Bk’!. Chem. 244:1314-24, l969. (Cited 230 timesj
3. GlIham PT. Synthesis of polynucleotide.ceUuloses and their use in fractionation ofpolynucleotiden. J.Amer. Chew. Soc.
86:4932-5, 1964. (Cited l9Otimen.)
4. Swan D, Aviv H & t.,eder 0. Purification and properties of biologically active messenger-RNA for a rnyeloma light ch~ia.
Proc. Not. ,lcad. Sd. 1/5.4 69:1967-71, l972. (Cited 245 limes.)
5. Roan .1, Aviv H, Scoinlek E & Leder P. In-vitro synthesis of DNA complementary to purified rabbit globin
messenger.RNA. Proc. Nat. ,jcad. Sc!. 03.4 69:264-8, 1972. (Cited 230 times.)
Received February 8,1991
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