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Authors: David Owen

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Julia Leigh's
The Hunter
(Penguin, 1999) became an international bestseller while attracting considerable flak. The storyline is as minimalist as its central character, M, a man with SAS-style hunting and survival techniques who is sent to Tasmania by a biotechnology corporation to find and kill a thylacine and take samples of its DNA for military purposes. He is successful in his mission. The fate of the thylacine? He buries it. (In a perverse real-life twist, word is out that some Tasmanian hunters say that if they discover and kill a thylacine they'll bury it and tell no-one, to prevent their shooting territory being locked away. Perhaps it's already happened! Lots of times!)

The novel was widely praised by UK and US critics, for its spare prose and philosophical overlay in the manner of a Moby Dick quest, pitting man against nature. It also drew comparisons with Ernest Hemingway's
The Old Man and the Sea.
But closer to home, and particularly in Tasmania, Sydney-based Leigh's use of the state was thought by some to be both gratuitous and inaccurate. It led to a debate, still unresolved, about the appropriation in fiction of a place with which an author is but passingly familiar. For University of Tasmania reviewer Andrew Peek, ‘
The Hunter's
setting, in rural Tasmania, pits hippie and conservationist against backwoodsy yokels. The bush is by turns impassive and threatening. In the best tradition of Tasmanian Gothic, it is a place of ghostly presences, beautiful, eerie landscapes and “devils” . . .'
14

Tasmanian discomfort with and dismissal of Leigh's novel became, in some instances, outright attack when Chloe Hooper's
A Child's Book of True Crime
(Random House, 2002) hit the bookstands. Like Leigh, Hooper is not Tasmanian—she lives in Melbourne—and her novel was also snapped up internationally (while still at manuscript stage by New York superagent Andrew Wylie) and published in great numbers to general acclaim. In this story Kate, a young teacher in a fictional town near Port Arthur, has an affair with the father of her precociously brightest pupil. His wife is the author of an acclaimed true-crime book on a foul local murder committed twenty years previously, which ominously involved a young mistress. Then strange, rather spooky things start to happen to Kate, and . . .

The thylacine element in this novel is satirical and tangential to the actual storyline, in the form of a parallel children's story in which a crime is investigated in the animal world by Terence Tiger, Kingsley Kookaburra and others (move over, Beth Roberts) but the tiger usage firmly roots the novel in its ‘Gothic Tasmanian' setting, as does its location in Port Arthur, scene of the 1996 slaying of thirty-five people by a deranged Hobart gunman. The front cover of the first hardback edition of the novel has an innocent, cartoon-style drawing of a group of Tasmanian animals, with Terence Tiger the thylacine prominent. How many copies his distinctive stripey image has sold will never be known.

14 BEATING
A SEVENTY
-YEAR HICCUP:
CLONING

My mother well remembers many years ago when her brothers brought home a young Tiger (they had caught the mother in a snare) & chained it in a corner of a room in the house for about a fortnight. They fed it on kangaroo and wallaby meat, & she says it used to growl & snarl if anyone went near while it was eating . . .

M
RS
O
LIVE
S
IMMONS,
S
COTTSDALE

W
hy clone a thylacine? The answer, in keeping with the contrary phenomenon of the animal, is both complex and divisive.

Artificial manipulation of flora and fauna to suit human requirements is an ancient practice, dating back to the domestication of wheat and subsequent selective cross-breeding for improved yields. The domestic dog has been interbred to achieve specific characteristics: tenacity, docility, endurance, speed. Chimeras—such as the liger, the offspring of a lion and tiger— while the product of natural reproduction, are unusual in that they are the product of two genetically different species. In all of this genes are the key.

A gene is a unit of deoxyribonucleic acid (DNA, sugary complex molecules) and strings of genes are arranged, or sequenced, on the chromosomes of a cell's nucleus. Genes pass on specific characteristics from parents to offspring. The full set of chromosomes of an individual is known as the genome. (The human genome has between 30 and 40 000 genes.) It follows that the ability to isolate and remove the genome from a plant or animal is the first step to artificially recreating that organism identically (cloning).

The second step in cloning is to ensure that the replicated genome is perfect. The third step is to grow it—in the case of animals, through the otherwise natural procedure of gestation in a (surrogate) mother.

This book will not have been too many years on library and home shelves when the Australian Museum's project to clone a thylacine is considered a success, a failure, or something else altogether. Begun in 1999, the project, with museum director Mike Archer at the helm, aims for a successful completion by 2010. In its early stages it has been characterised by both scientific breakthroughs and scientific and ethical problems.

Archer is a conservation biologist with a particular interest in extinct species. His PhD was on Australia's carnivorous marsupials and he has since done much in the palaeontological line, including recent discoveries of seven ancestral thylacine fossils in Queensland, ‘in about four genera [closely related species], only one of which is
Thylacinus
'.
1
His credentials as a thylacine expert and devotee are unquestioned. When he came across a preserved thylacine pup in the Museum's collection many years ago, he paid it more than passing attention, later reasoning that because it had been kept in ethyl alcohol rather than formalin (since 1866), its DNA might not have been destroyed and therefore— at some future point—might successfully be extracted.

Seed funding for this supremely ambitious project was provided in 1999 by the New South Wales government and a private trust. Obtaining money was in all likelihood the easiest part of the equation. Certainly, the pitch for funding must have sounded good: ‘What Mike Archer and his team are attempting is as scientifically exciting and technically challenging as splitting the atom or landing a man on the moon'.
2

Significantly trickier was the question of the DNA itself: acquiring it from a variety of sources, successfully extracting it, hoping that it would be undamaged, replicating it. Thylacine remnants are scattered in museums and other collections around the world, legacies of the sixty-plus animals known to have been displayed in zoos between 1856 and 1936. Obtaining access to such rare and valuable material, for the sake of genetic variety, would prove crucial. Did the Australian Museum have access to it? ‘Yes. Most of this knowledge about other specimens has been coordinated via the Tasmanian Museum and Art Gallery in Hobart.'
3

Thylacine DNA is aged, fragmented and dehydrated. Its reconstruction is ferociously complicated, even when explained simply:

The ethyl alcohol in which infant thylacines have been stored preserves tissues and genes, but in doing so, water, the very substance which is vital to life, is expelled. Scientists would need to rehydrate not only the DNA in the thylacine nuclei, but the entire matrix of nuclear proteins and enzymes that maintains its structure, and oversees its orderly replication as the cell divides and multiplies to form an organism. Therefore, more is needed than just the coded instructions for how to build a thylacine. Obtaining the complete package of necessary biochemical elements will be the determining factor in whether thylacine cloning is a success or failure.
4

The Australian Museum team set to work and DNA—not without difficulty—was isolated and extracted from the 1866 pup and subsequently from two other pups. The tissue sources included bone, bone marrow and muscle. The extracted DNA, however, represents only a miniscule percentage of the complete genome of the specimens.

Three years into the project a significant breakthrough was announced. Through a process known as polymerase chain reaction (PCR), four thylacine genes were successfully replicated and copied into millions of short fragments of undamaged DNA.

That is where the project apparently stood in late 2002, with the next stage to involve the construction of synthetic chromosomes from the fragments, building towards the perfectly replicated genome. Archer refers to it as creating a ‘genetic library' of a thylacine's DNA. (The metaphor is apt. The Library of Congress has holdings of many millions of items, each one with a specific classification assigning it to one and only one place on a shelf.) How is the library actually built?

The concept is that the DNA of the thylacine is tagged onto the DNA of living creatures (plasmids of yeast or bacteria) which then look after the DNA—and this is the library. We are recovering DNA from three individuals. Whether that means each will have a library of its own, I couldn't tell you. This is an issue that the geneticists are better able to answer.
5

Professor Marilyn Renfree has gone some way towards providing an answer. In February 2002 she was one of four experts on an edition of the ABC-TV science programme,
Catalyst
, looking at the thylacine cloning issue. Renfree, who teaches at the University of Melbourne's Department of Zoology and is a specialist in marsupial reproduction, cited the current difficulties in cloning with living, undamaged material. Thus, Dolly the cloned Scottish sheep, while an undoubted scientific first, had malformed organs and died suddenly in 2003. For Renfree, cloning is ‘extraordinarily difficult . . . Dolly was one egg in 277 fertilisations, so there were 277 nuclei put into 277 enucleated eggs and we got one Dolly'.
6

On that same television programme, Dr Ian Gunn of the Animal Gene Storage Institute of Australia was even more sceptical: ‘To construct a nucleus and the DNA is virtually asking someone to construct a new species. To me, impossible'.
7

A counter-argument is that science of this type can but improve. Another is that ‘big science'—meaning a big story— attracts money that otherwise wouldn't go the way of science. And Archer himself pulls no punches: ‘Personally, I think this is the most exciting biological project that's going to occur in this millennium'.
8

Use of a host to build this library apparently poses no problems: ‘If a thylacine is reconstructed and brought to life through this process, using a surrogate such as a Tasmanian devil, it will be a thylacine—and nothing but. With only thylacine nuclear DNA to build the animal from, it can't be anything else'.
9

Even if that is so, the required use of a host presents problems. To date, geneticists have not been able to grow nuclei taken from one living animal in another that is related, because the host oocyte (female germ cell) consistently rejects the implanted nuclei.

It would be a miraculous birth, a clone from aged DNA. It would, no doubt, ‘be' a thylacine physically. But would it have an inherited ‘personality' from the lineage of the 1866 pup? Or, like a captive-bred animal, might it lack a range of acquired behavioural traits as learned when wild-raised?

This is a bit of suck it and see. I am convinced that a lot (most) marsupial behaviour is ‘hard-wired' and under the control of the genome. This is the reason that behaviours of individuals raised in captivity commonly closely mimic the behaviour of individuals studied in the wild. But, at this point, this is speculation.
10

Ultimately it is proposed that a minimum number of thylacines would be cloned, and that natural reproduction would then take place followed by release into a safe area. ‘No-one is clear yet about where the best place would be. We'll be leaving that up to the Tasmanian Parks and Wildlife Service, if the project is successful.'
11

It's worth remembering that in 1967 the Fauna Board under Eric Guiler selected Maria Island as a sanctuary, should thylacines ever be found. The uninhabited island still holds that status, a fact of which Col Bailey reminded his audience when he gave a talk at the Tasmanian Museum and Art Gallery in September 2001.

Research into cloning extinct and endangered fauna has been going on elsewhere, and meeting with criticism as well as support. In Australia, the Mike Archer-led thylacine project has met with five main areas of criticism.

Many within and beyond the scientific community doubt that it can be done at all:

While managing to get PCR to work on a degraded source of DNA like a preserved pup is commendable, this breakthrough represents the tip of the iceberg in terms of creating a living, breathing thylacine . . . The ‘hurdles' of decoding the correct sequence of DNA, constructing synthetic chromosomes and transferring chromosomes into a cell are tasks of such mind-numbing complexity as to make the PCR amplification of a few genes look like child's play.
12

The cost of the project, already high and still rising, has inevitably led to complaints that research efforts could be better spent elsewhere, not least in slowing Australia's alarming rate of extinctions. There is also scepticism at the proposal to repopulate Tasmania's wilderness, given that each cloned thylacine may be worth millions of dollars. The need to protect them and the financial windfall to be had through displaying them suggests, instead, a form of captivity.

Captive thylacines did not breed, despite instances of males and females being kept together. And despite the optimism of some, such as Ronald C. Gunn, superintendent of convicts at Launceston, who in 1850 wrote of the pair destined for the Royal Zoological Society, ‘I feel little doubt but that the Thylacines will do well and very probably breed . . . I can imagine nothing in the climate of London likely to injure them very materially'.
13
The chance of clones breeding naturally in captivity cannot be rated highly. That would suggest the need for articifial insemination (as is so often the case with the panda), removing further the reconstituted animal's naturalness.

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