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JCVI Scientists Successfully Transplant Bacterial Genome

28 June 2007

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Colonies of the transformed Mycoplasma mycoides bacterium. Click to enlarge.

Researchers at the J. Craig Venter Institute (JCVI) have successfully transplanted the genome from one species of bacterium into another.

The work, published online in the journal Science, by JCVI’s Carole Lartigue and colleagues, outlines the methods and techniques used to change one bacterial species, Mycoplasma capricolum, into another, Mycoplasma mycoides large colony (LC), by replacing the genome of the former with that of the latter.

The successful completion of this research is important because it is one of the key proof of principles in synthetic genomics that will allow us to realize the ultimate goal of creating a synthetic organism. We are committed to this research as we believe that synthetic genomics holds great promise in helping to solve issues like climate change and in developing new sources of energy.

—J. Craig Venter, Ph.D., president and chairman, JCVI

Genome transplantation is an essential enabling step in the field of synthetic biology as it is a key mechanism by which chemically synthesized chromosomes can be activated into viable living cells. The ability to transfer the naked DNA isolated from one species into a second microbial species paves the way for next experiments to transplant a fully synthetic bacterial chromosome into a living organism and if successful, “boot up” the new entity, according to JCVI scientists.

The JCVI team devised several key steps to enable the genome transplantation. First, an antibiotic selectable marker gene was added to the M. mycoides LC chromosome to allow for selection of living cells containing the transplanted chromosome.

Then the team purified the DNA or chromosome from M. mycoides LC so that it was free from proteins (“naked DNA”). This M. mycoides LC chromosome was then transplanted into the M. capricolum cells. After several rounds of cell division, the recipient M. capricolum chromosome disappeared having been replaced by the donor M. mycoides LC chromosome, and the M. capricolum cells took on all the phenotypic characteristics of M. mycoides LC cells.

As a test of the success of the genome transplantation, the team used two methods—2D gel electrophoresis and protein sequencing—to prove that all the expressed proteins were now the ones coded for by the M. mycoides LC chromosome.

Two sets of antibodies that bound specifically to cell surface proteins from each cell were reacted with transplant cells, to demonstrate that the membrane proteins switch to those dictated by the transplanted chromosome not the recipient cell chromosome. The new, transformed organisms show up as bright blue colonies in images of blots probed with M. mycoides LC specific antibody.

The group chose to work with these species of mycoplasmas for several reasons: the small genomes of these organisms which make them easier to work with, their lack of cell walls, and the team’s experience and expertise with mycoplasmas. The mycoplasmas used in the transplantation experiment are also relatively fast growing, allowing the team to ascertain success of the transplantation sooner than with other species of mycoplasmas.

While we are excited by the results of our research, we are continuing to perfect and refine our techniques and methods as we move to the next phases and prepare to develop a fully synthetic chromosome.

—Dr. Carole Lartigue

This research was funded by Synthetic Genomics Inc. Synthetic Genomics, which was founded by Venter, recently entered a research and development partnership with BP focused first on gaining a better understanding of the natural microbial communities in various hydrocarbon formations such as oil, natural gas, coal and shale. The next step in the partnership is then to develop organisms optimized for the enhancement or increased production of subsurface hydrocarbons. (Earlier post.)

Venter is also one of the signatories of the Ilulissat Statement, which calls for an international effort to advance synthetic biology that would not only propel research, but do so while developing protective measures against accidents and abuses of synthetic biology. (Earlier post.)

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June 28, 2007 in Biotech, Fuels, Synthetic Biology | Permalink | Comments (7) | TrackBack (0)

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Comments

Welcome to the biorobotic era of living machines!
The dream synthetic organism would take in basic material like cellulose, sunlight and CO2, or even electricity and CO2 and then putout ethanol, butanol, petroleum, plastic monomers, pharmaceuticals, anything you want in highly pure amounts with no waste.
Of course another dream of synthetic biology is to making superhuman bioroids that will do your bidding, but that’s another story.

Cool!


FRANKENBACTERIA !!!

I'm not getting all excited again. When they cloned "Dolly" the sheep, I thought I would get to have my own copy of Catherine Zeta Jones, and so far, no dice.

For bacteria, I think transplanting all the DNA is less interesting than planting the select bits. I assume that's what the biotech companies are doing combining jungle rot bacteria with some genes from yeast, but high concentration ethanol kind of kills bacteria, although maybe if the combine it with some archaea extremophiles from a sulfur vent or something.

We will still be transplanting bits and pieces for some time, synthetic biology will eventually provide a more efficient and controllable platform then genetically modified organisms, aka the "bioroid". Say for example we could with genetic modification get bacteria that can withstand maybe 10-15% ethanol concentrations, but with organism completely built from scratch with every gene made to produce ethanol optimized proteins and enzymes we could do maybe 90%! Of course to do that we would need quantum computers to design and optimize enzymes to that level because predicting protein folding and shape with high accuracy requires herculean calculations.

This article by Venter and colleagues is a very tiny forgettable technical feat in gene transfer. The hype is many years behind serious synthetic biology. It is as if people who build cars that use ethanol-containing fuel start claiming that they have revolutionized transportation. Nope. Not yet.

or like Otto Lilienthal claiming power flight will be the future after testing his first glider.

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