The genetic code

As it became evident that genes cotrolled the structure of polypeptides attention focused on
how the sequence of the four base pairs in DNA could control the sequence of the 20 amino acids
found in proteins. With the discovery of the mRNA intermediately, the question became one of how
the sequence of the four bases presents in mRNA molecules could specify the amino acids sequence
of a polypeptide. What is the nature of the genetic code relating mRNA base sequences? Clearly,
the symbols or “letters” used in the code must be the bases, but what comprises a codon, the unit or
“word” specifying one amino acid (or actually one aminoancyl-tRNA complex) shown (by backcrosses
to wild type : fig 11.9) to result than from back mutation at the original site of mutation. Crick and
colleagues reasoned that in the oraginal mutation wase a sigle base-pair delection or addition,
respectively, occurring at a site near the original mutation. A sigle base-pair addition or delection will
alter the reading frame of the gene and mRNA (the codons in hase during translation) for that
portion or the gene distal to the mutation (relative to the direction of translation) this is illustration
in fig 10.30a when the supressor mutation were isoleted or sigle mutans by screening progeny and
backccrosses to wild type, they were found to produce mutant phenotypes, just like the original
mutation. Crick and colleague next isolated provalin induced suppressor mutations of the original
suppressor mutations, and so on

All the isolated mutations were then classified into two groups, plus (+) and minus (-) (from
addition and delection all thought Crick et all has no idea which groups was which) using the
reasoning that a (+) mutation would suppress a (-) mutation, but not another (+) mutations and
which versa (gambar 10.30) and legent for additional legent) nexts Crick et all contructed
recombinants that carried various combinasionals of the (+) and the (-) mutationts. Recombinans
with two (+) mutationts or two (-) mutations always had mutans phenotypes, just like the single.

Three Nucleotides per codo

Twenty diferents amino acids incoroarated during translation. Thus at least 20 different
codons must be formed using the four symbols (bases) available in the “messanger” (mRNA). Two
bases per codon would yield only 43 or 64 possible codons-an apparent excess.

The first strong edivence that the genetic code was in fact a triplet code (Three nucleotides
per codon) resulted from a genetic analysis of provalin-induced mutations in the rill locus of phage
T4 carried out by F. H. C. Crick and colleagues in 1961. Crick and coleagues isolated proflavin-induced
revertaints of a proflavin-induced mutation. (prilavin, an acridine dye induces singles base pair
additions and deletions see chapter 11 pp 310-311) these revertains were

Figure 10.30 (right pase) schematic illustration of crick and coworker’s proof that the genetic
code is a triplet code (three bases per codon). Crick and colleagues studied a series of supressed
mutation of a mutation at the rill locus of phage T4. The original rII mutation had been induced with
result of a sigle base-pair addition or delection. In (a) the original mutation is shown arbitarily as a
sigle base-pair addtion, specifically as an AT base-pair insertion (wild type allele mutant allele) the
nucleotide pair sequence shown for the wild type allele (and thus teh mRNA base sequence and
amino acid sequence of the polypeptide) is hypothetical crick and coworkers selection phenotypic
revertants of this mutant and demostrated by backcrosses that these revertans resulted from
suppressor mutations, not back mutations at the original mutans site. If the original mutation is an
addition gene product activity might be restrored by a delection (single base-pair) mutation in a
nearby region for example the delection of a CG base-pair as shown (mutants allele revertaints
allele) the original addition mutation will change the reading frame (determining codons in phase for
translation) for all codons distal (relative to the direction of translation) to the site of the mutation.
The subsequens delection (suppressor mutans) will restore the reading frame to the distal portation
of the gene. If the altered amino acid sequence is not critical to runction the protein produced by the
double muatnt gene will be active. When suppressor mutations were isolated in single muatans
strains by backcrosses to wild type, all there found to yield muatns phenotypes just like the original
mutations that they supressed. Crick and colleague then the isolated proflavin induced suppressor
mutation of the proviously isolated suppressor mutations (present in a single mutans recovered
from backcossed). After repiating this procces for several cycle, all the mutations were classifield as
plus (for sigle base-pair addition) or minus (for single base-pair delection) on the basis taht a plus
mutation would suppress a minus mutation but not another plus mutation, and vice versa actually
crick and colleagues had no idea wheter the plus group of mutations represented addition or not
they could just as likely have been delectio. The only importaints point was that all delection ended
up in one groups (be it the plus group or the minus group) and all addition ended iup in the other
group.

Having so classifield the mutations, crick and cowokers next performed the critical
experiment. They isolated recombinats carryng various combination of plus and minus mutations.
When two plus mutations were present in a recombinats its phenotype was always mutans. The
same was true in the case of the recombinations. These result are most easily explained if each
codon contains three bases mutans. Recombinants carryng three (+) mutation or three (-) mutation,
however often had wild type phenotypes. This indicated that the addition of three base pair or the
delections of tree base pair left distal portains of the gene with the correct (wild type) reading frame
a result that would be expected only if each codon contained three nucleatides