Molecular biology does much the same. First, it needs to multiply the number of copies of the message to allow each short piece to be surveyed in detail as a preliminary to the complete map. Various tricks allow cut pieces of DNA to be inserted into that of a bacterium or yeast. The DNA has been cloned. Whenever the host divides, it multiplies not only its own genetic message but the foreign gene. As a result, millions of copies of an original are ready for study in the exquisite detail needed for genetic geography.
Cloning has been supplemented by another contrivance, the polymerase chain reaction. This takes advantage of an enzyme used in the natural replication of DNA to make replicas of the molecule in the laboratory. To pursue our rather tortured literary analogy, the method is a biological photocopier which can produce many duplicates of each page in the genetic manual. The photocopying enzyme comes from a bacterium which lives in hot springs. The reaction is started with a pair of short artificial DNA sequences which bind to the natural DNA on either side of the length to be amplified. By heating and cooling the reaction mixture and feeding it with a supply of the four bases, the targeted strands of DNA unwind, copy themselves with the help of the enzyme, and re-form. Each time the cycle is repeated, the number of copies doubles and millions of replicas of the original piece of DNA are soon generated.
Another piece of trickery exploits DNA’s ability to bind to a mirror image of itself. DNA bases form two matched pairs; A with T and G with C. To find a gene, a complementary copy is made in the laboratory. When added to a cell this seeks out and binds to its equivalent on the chromosome. My computer can do the same. On a simple command, it will search for any word I choose and highlight it in an attractive purple. It does the job best with rare words (like ‘banana’). A DNA probe labelled with a fluorescent dye shows up genes in the same way. The method is known as FISHing (for Fluorescent In-Situ Hybridisation) for genes. A modified kind of FISH involves unwinding the DNA before it is stained. This makes the method more sensitive.
All this and much more has revolutionised the mapping of human DNA. First, it has improved the linkage map. Patterns of inheritance of short sequences of DNA can be tracked through the generations just as well as can those of colour-blindness or stubby fingers. There are millions of sites which vary from person to person. All can be used in pedigree studies. Another scheme is to use the polymerase chain reaction to multiply copies of DNA from single sperm cells. The linkage map is made from a comparison of the reordered chromosomes in the sperm with that in the man who made them. This avoids the problem of family size altogether.
Linkage mapping in humans took a long time to get started and still has some way to go. Before the days of high technology the great problem was a shortage of differences; of variable genes, or segments of genetic material, whose joint patterns of inheritance could be studied. That problem has been solved. Our DNA is now known to be saturated with hundreds of thousands of variable sites, many based on individual variation in the numbers and positions of repeats of the two letters C and A. As a result, a whole new industry based on the most traditional kind of genetics has burst into existence.
It needs, like any industry, raw material. The French, together with the Americans, have identified sixty or so large families with long and complicated pedigrees, well suited for gene mapping. They come from various parts of the world, from Venezuela to Bangladesh. From each individual, lines of cells are kept alive in the laboratory and thousands of variants have been identified, tightly packed along the entire length of the chromosomes. Patients with, say, heart disease can be screened to see whether they also tend to carry other inherited variants. If they do, there is a good chance that the actual gene involved is nearby, and is dragging its anonymous fellows along with it. To find such a milestone may be the first step to the gene itself.
The descendants of Morgan have at last managed to do for humans what was long ago achieved for the fruit fly, and a linkage map of man is close at hand. That of woman, it transpires, is rather longer. Such maps depend on the sexual reshuffling of genes. This takes place, for some reason, more in females than in males and, as a result, their chart works to a different scale.
The human linkage map is useful, but biologists have always wanted to make a different kind of chart, one rather like that used by geographers, based on a straightforward description of the genetic material. Now, it is here. The approach was brutal: to assault the genome with time, money and tedium until the whole lot was read from one end to the other.
The first move in tying the linkage map to one based on the physical structure of DNA depended on a stroke of luck. Morgan noticed that in one of his fly stocks a gene which was usually sex-linked started behaving as if it was not on the X chromosome at all. A glance down the microscope showed why. The X was stuck to one of the other chromosomes and was inherited with it. A change in the linkage relationships of the gene was due to a shift in its physical position.
Such chromosomal accidents were used to begin the human physical map. Sometimes, because of a mistake in the formation of sperm or egg, part of a chromosome shifts to a new home. Any parallel change in the pattern of inheritance of a particular gene shows where it must be. Now and again a tiny segment of chromosome is absent. That can lead to several inborn diseases at once. One unfortunate American boy had a deficiency of the immune system, a form of inherited blindness, and muscular dystrophy. A minute section of his X chromosome had been deleted. It must have included the length of DNA which carried these genes. He gave a vital hint as to just where the gene for muscular dystrophy – one of the most frequent and most distressing of all inherited diseases – was located. The absent segment was a landmark upon which a physical map of the area around this gene could be anchored.
To map genes with changes in chromosomes need not wait for natural accidents. Human cells, or those of mice or hamsters, can be cultured in the laboratory. When mixtures of mouse and human cells are grown together, the cells may fuse to give a hybrid with chromosomes from both species. As the hybrids divide, they lose the chromosomes (and the genes) from one species or the other. Some specifically human genes are lost each time a human chromosome is ejected. To match the loss of particular genes with that of chromosomes (or of their short segments) shows where they must be.
All these methods hint at a gene’s position rather than giving its precise coordinates. Small-scale cartography (or mindless sequencing, as it is affectionately known) involves various clever ruses. One depends on the ability of DNA to copy itself when a special enzyme is provided and the mixture fed with the A, G, C and T bases. It is possible to gradually lengthen pieces of a DNA strand from one end to the other, in four separate experiments (each using a different base). By chemical trickery, some of the growing strands are stopped each time a base is added. This produces a set of DNA pieces of different length, each stopped at an A, a G, a C or a T. Electrophoresis of the mixtures on the same gel gives four parallel lines of DNA fragments arranged by length. A scan across and down the gel gives the order of the bases. This is a most tedious task. It has been supplanted by machines that do the job in other ways. The most important change in genetics is a conceptual one. Because the three-letter code for each amino acid is known, it is possible to deduce the order of the amino acids made by a piece of the DNA once its sequence of bases has been established. What any gene does can be inferred by comparing that sequence with the computer database of others whose job is known. The fit need not be precise; after all, a French dictionary contains thousands of words similar enough to those in English to allow its meaning to be guessed at. It is also sometimes possible to work out the three-dimensional structure of the protein from its amino acid sequence and to deduce what its function might be.
There are some remarkable similarities among inherited vocabularies. The genes that control development are similar in humans and fruit flies, as are those that make their brains. Genes that, when they go wrong, damage the nervous system have close analogues in yeast (which do not have nerves at all) and one of our own genes is almost identical to another that alters the pattern of veins on an insect wing. Such conservatism has had a radical influence on human genetics.
The parts catalogue for a Mercedes C-class car contains four and a half thousand named items, from accelerator pedal