Plant Protein Club biol46 at
Mon Nov 2 21:10:28 EST 1998


Transient Expression in Plant Protoplasts: A rapid screening method for testing activity and processing of recombinant proteins 
in plants

22 - 25 March 1999, University of York, UK

Transient expression of plasmid-borne genes in plant protoplasts is a powerful technique to study gene expression at all levels. 
An important advantage of this technique is its speed compared to the tedious stable transformation methods. Results can be 
obtained in days rather than the months usually required for regeneration of transgenic plants, because gene products are 
detectable within hours after transfection. This technique has been used successfuly to study regulatory domains of promoter 
regions, mostly making use of reporter fusions allowing simple enzyme assays. 

More recently, transient expression has been used for other purposes, such as testing the activity, processing and targeting of a 
recombinant protein. The method is also ideal to study transcript maturation or to test mRNA stability or the efficiency of 
translation initiation. Analysis of protein stability, activity, transport, post-translational modification, folding and oliglomerization is 
also possible, but these refined methods require routine preparation of large numbers of protoplasts. 

Course aims

The principle aim of this course is to introduce scientists from academia and industry to this technique and its various 
applications. The emphasis will be on the practical aspects, and participants will be able to carry out the technique step by step 
under the supervision of scientists who use this method every day. In addition, discussion sessions will address troubleshooting, 
optimisation, and tricks and traps, with the objective of assisting those who are already using the technique. Specialised 
applications as well as routine problems will be addressed with equal emphasis. 

Course tutor

Dr Jurgen Denecke is a lecturer at the University of York (UK). He has extensive experience in developing and utilising the 
technique of transient expression in plant protoplasts. His interests and current research include protein synthesis, folding and 
transport in the endoplasmic reticulum and plant biotechnology. 

Course programme

The course programme will be composed of practical work and lectures. A range of techniques will be demonstrated and 
taught. There will be opportunities for participants to discuss their own projects and to resolve their specific problems in relation 
to this technique. 

Examples of techniques taught include: 

· protoplast preparation 
· electroporation of protoplasts 
· protein gel eletrophoresis 
· protein gel blotting 

The course will also include experiments that demonstrate the use of reporter proteins and teh combination of subcellular 
fractionation with transient expression to study protein targeting. Other techniques will be discussed theoretically. During the 
course, time will be allocated to allow participants to test their own plasmids, although you may have to bring the necessary 
reagents for downstream analysis and should contact PPC for advice. The content of the course is tailored to meet the specific 
requirements and backgrounds of participants. To this end, all participants are asked to complete a pre-course questionnaire. 
Course registration and fees information

INDUSTRIAL RATE: £550 (PPC members £350)
ACADEMIC RATE: £325 (PPC members £200)

The registration fee covers tuition, course handbook, a comprehensive collection of protocols for further reading, experimental 
materials as specified within the course programme, lunch and refreshment breaks each day. Course fees do not cover 
specialised chemicals required for downstream analysis of participants' own plasmids. Accommodation is not included. 

For a registration form and pre-course questionnaire or further details, contact PPC.

Plant Protein Club
PO Box 373
University of York
York YO10 5YW

Tel/Fax: +44 (0)1904 434327
Email: ppc at
WWW site:

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