Before the rise of molecular biology in the 1950s and 1960s, a small number of biologists had explored the possibilities of using biochemical differences between species to study evolution. Alfred Sturtevant predicted the existence of chromosomal inversions in 1921 and with Dobzhansky constructed one of the first molecular phylogenies on 17 Drosophila Pseudo-obscura strains from the accumulation of chromosomal inversions observed from the hybridization of polyten chromosomes. Ernest Baldwin worked extensively on comparative biochemistry beginning in the 1930s, and Marcel Florkin pioneered techniques for constructing phylogenies based on molecular and biochemical characters in the 1940s. However, it was not until the 1950s that biologists developed techniques for producing biochemical data for the quantitative study of molecular evolution.
The first molecular systematics research was based on immunological assays and protein "fingerprinting" methods. Alan Boyden—building on immunological methods of G. H. F. Nuttall—developed new techniques beginning in 1954, and in the early 1960s Curtis Williams and Morris Goodman used immunological comparisons to study primate phylogeny. Others, such as Linus Pauling and his students, applied newly developed combinations of electrophoresis and paper chromatography to proteins subject to partial digestion by digestive enzymes to create unique two-dimensional patterns, allowing fine-grained comparisons of homologous proteins.
Beginning in the 1950s, a few naturalists also experimented with molecular approaches—notably Ernst Mayr and Charles Sibley. While Mayr quickly soured on paper chromatography, Sibley successfully applied electrophoresis to egg-white proteins to sort out problems in bird taxonomy, soon supplemented that with DNA hybridization techniques—the beginning of a long career built on molecular systematics.
While such early biochemical techniques found grudging acceptance in the evolutionary biology community, for the most part they did not impact the main theoretical problems of evolution and population genetics. This would change as molecular biology shed more light on the physical and chemical nature of genes.
Read more about this topic: History Of Molecular Evolution
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