Catching up with Mark

By Mia Cerfonteyn

At The Sainsbury Laboratory, we try to understand how different cellular components function as part of the plant’s immune system. To test our hypotheses, we often need to be able to remove, add or modify proteins within biological systems. This is why we need people like Mark Youles and his colleague, Liam Egan, who make up our Synthetic Biology division.

“The name Synthetic Biology might be a little misleading for our group.” Mark admits when I met with him on a summer’s day outside The Sainsbury Laboratory to learn more about what his Synthetic Biology (SynBio) group actually does.

Mark explained that the TSL Synbio group primarily supports The Sainsbury Laboratory in their synthetic approaches to complex cloning issues (which is akin to cutting and pasting bits of DNA). This is aimed at the biological engineering of plants to produce higher yields, endure environmental stress, and increase disease resistance. The Synbio service also provides the wider Norwich Research Park with a molecular library of cloning parts which enable research teams to achieve their goals.

The correct assembly of golden-gate constructs can be monitored by the production of coloured proteins in their bacterial hosts.

Golden Gate cloning method

The original Golden Gate method, published by Sylvestre Marillonnet and colleagues in 2009, outlined the extremely efficient and accurate assembly of multiple DNA fragments in a predefined order into fully assembled constructs.

The defining characteristic of the Golden Gate method is its use of alternating pairs of type IIs restriction enzymes which allow many individual DNA fragments to be cut and pasted together within a single ‘one pot’ reaction, saving researchers substantial amounts of time and labour.

An example of Golden Gate cloning allowing three fragments to be ligated in a specified order in one step.

Working as a technician

To fully understand the complexity of these systems requires an intimate knowledge and an advanced level of expertise of all of the processes and methods involved. Luckily for us, Mark has had over 20 years of experience in this and closely related cloning fields.

Mark comments, “I’m lucky that I have been able to see these methods evolve throughout my career. While cloning was sometimes a tedious and unpredictable process when I started out, that experience has significantly helped me to fully understand and appreciate the mechanisms behind what we’re trying to achieve today. This understanding is crucial for identifying and solving any problems which occur in our experiments, especially in the fast-paced research environment of TSL.”

Mark Youles in the laboratory. Photo by Steph Bornemann.

With the rapid advancement of technologies in fields such as Synthetic Biology, Mark is concerned that we are facing a decline in the fundamental knowledge of many earlier cloning methods.

“It’s important that experienced support staff have the opportunity to transfer their skills and knowledge to those who will supersede us.” Mark says, “There are currently no formal courses that can teach someone what I know. Most of the intricate methods and techniques need to be passed on by personal tuition."

"Although most university institutions teach the established principles of general cloning, these are being superseded by faster modern kits and techniques. Ultimately, this dilutes the critical and fundamental understanding of many of the important elements and processes involved. Golden-gate cloning is a classic example of such a technical advancement, but still requires a full understanding of what is happening at each and every step of the process.”

Successful research institutes, such as The Sainsbury Laboratory, depend on a strong foundation of driven and experienced support staff. Our dedicated staff are the reason the Laboratory has been able to consistently produce research of excellent quality and push the boundaries of our knowledge on plant microbe interactions.

“I’m very proud of all of the pipelines and capacity we’ve been able to build here at The Sainsbury Laboratory." says Mark, "Our experience with the golden-gate system and cloning in general has spread across the Norwich Research Park, meaning we are now also often asked to provide cloning information and advice to many of the other institutes located throughout the whole of the Park.”

Butterflies and their secrets

As we look out at the flurry of pollinators hovering around the meadow flowers in the courtyard, Mark tells me about his fascination with butterflies and moths which stretches back to his early childhood.

“I had one of those old-fashioned childhoods where I would leave the house in the morning and only return later that evening.” Mark reflects, “Moths and butterflies provided me with endless hours of outdoor interest, fascination and entertainment. I spent much of my childhood collecting and rearing caterpillars with an ultimate aim of attempting to breed them.

European Purple Emperor butterfly (Apatura ilia). Photo by Mark Youles.

After training as a scientist, Mark was able to look at his childhood hobby through very different eyes.

“There are still so many crucial, unanswered questions about the Purple Emperor butterfly”, Mark informs me is his primary butterfly of interest.

“Back in 1857, it was first reported that males of these butterflies were attracted to carcasses and dung, but even 164 years later nobody knew why they did this." he says, "Since starting my research, I have identified that like many other species of butterfly, the male Purple Emperor’s possess male specific pheromones."

Deuterium labelling experiments conducted by a collaborator defined that branched chain amino acids (predominantly valine) sequestered from the dung and carcasses form a principle component that is incorporated directly into the male specific pheromone with very high efficiency.

Last year Mark also supplied an additional collaborator with specimens which were used to extensively analyse the visual capacity of the eyes of both male and female specimens.

He goes on to explain, “We were looking for any kind of sexual dimorphism, but despite very careful analysis we ultimately concluded that there wasn’t any significant difference in the visual capacity of either sex.”

Mark highlights the alarming rate at which many of Britain’s butterflies are declining due to increasing conservational threat and habitat loss. “By fully understanding the factors which are essential to the successful breeding of these (and other) butterflies, we can ensure that such factors are successfully maintained within their natural breeding habitats."

Swallowtail butterfly (Papilio machaon). Photo by Steph Bornemann

Plant health and our future

Whether it is people, butterflies, or plants, Mark has always felt strongly about the purpose of his work.

“I need to believe in the goal of what I’m doing, otherwise I quickly lose motivation. This is probably why I spent the bulk of my early career supporting research for cancer therapies at the Medical Research Council in Cambridge.” he says.

Mark also loves inspiring younger generations to get excited about science. In this photo Mark and Liam are using lollipops to explain DNA structure at the Plants of the Future event held at Norwich Research Park in May 2022. Photo by Steph Bornemann.

I ask Mark what motivates him to come to The Sainsbury Laboratory every morning.

“I never really considered the future of our plants and our food supplies before I started working here.” he answers, “I openly admit that I didn’t initially see that plant diseases were anything that posed a significant threat. As global population numbers however continue to climb, it becomes increasingly obvious that our struggle to feed these exploding populations is likely to become a very real and difficult issue in the not too-distant future. We urgently need to get ahead of the curve regarding the important issues that affect plant disease resistance and increased crop yields in order to prevent future food shortage concerns.”

“Knowing what I know now, I feel very proud of the work we do here and am driven to go the extra mile in helping our researchers succeed in their projects.”

About life

As challenging as scientific research can be, life outside of work has a tendency to throw even bigger challenges our way.

Mark suffered a sudden brain haemorrhage on Southwold beach in July of 2021, and it has understandedly taken him some time to regain his strength and fitness. He has thankfully now made a full recovery and was finally able to return back to working here at The Sainsbury Laboratory in a full-time capacity towards the end of February 2022.

Mark shares that the incident made a very significant impact on his attitudes and approaches towards many things in life.

“Such a significant and sudden health scare really brings many things into focus. On initial assessment I was given around 1:3 chance of leaving the beach alive and it was completely unknown to what extent I could have incurred complete or lasting cognitive impairment. Making a full recovery so completely and quickly is a real lasting testament of how professional, talented and truly magnificent all of our emergency services here in the UK really are.”

Speaking a little further about his experience Mark also adds that, “It is really only during these times of crisis do we also really begin at appreciate how wonderful employers such as The Sainsbury Laboratory really are. They gave me all of the care and support that I could ever wish for. They simply could not do enough to ensure that my recovery could proceed at a pace that I was comfortable with. I truly felt fully supported at every stage, which just goes to show how much they really care, appreciate and look after their staff.”

Mark meets with the excellent EAAA and SARS emergency services that played such a critical role in saving his life.

Mark continues to actively support the East Anglian Air Ambulance (EAAA) in any way he can. He has helped to promote the EAAA lottery which provides the service with almost one third of their annual £15m annual operational donation funding and has also taken part in a documentary film which illustrates the importance of the EAAA aftercare support network.

Everyone at The Sainsbury Laboratory is very grateful for the excellent emergency services that took such great care of our deeply valued colleague and friend.

EAAA and TernTV produced this story about Mark's incredible recovery.