No subject

[Al Ivens]

Dear Colleagues,

This is a general message to let interested parties know that I am currently 
involved in a genome analysis project of Leishmania major Friedlin.  My 
background is the Human Genome Project (Heding et al. Genomics 13:89-94; 
1992), but I have moved over to the much more interesting (!) Leishmania.  I 
am particularly interested in Chromosome 14, which, along with my 
co-researcher Debbie Smith, has an interesting example of 
differentially-regulated gene expression (Flinn & Smith, Nuc. Ac. Res. 
20:755-762; 1992).

The aim of our project is to construct a cosmid contig map of the chromosome 
(approximately 900kb) and also to identify sequence tagged sites and/or 
expressed sequence tags.  These will hopefully be of use for comparative 
mapping purposes.

We have a L. major cosmid library in cLHYG.  A number of different research 
groups contributed to its construction, but I have now picked ~ 10,000 
individual clones into 96 96-well microtitre plates, and gridded them out in 
4x4 arrays on nylon membranes.  These have been hybridised with pulsed-field 
gel-purified L. major chromosomes to identify chromosome-specific cosmids.  
The technique appears to have worked pretty well, and we have identified 900 
clones specific (potentially) for chromosome 14.  A number of the smaller 
chromosomes have also been used as probes with similarly useful data being 
obtained.

I have just finished fingerprinting the clones, and am now in the process of 
putting together an overlapping clone map (contig), using Sun 
Workstation-based analysis software.  While I do not expect to get full 
coverage (I know the telomeres are missing from the library), I believe that 
a high degree of coverage will be obtained.  So much data to go through!

A number of other parallel studies, aimed at identifying more biologically 
interesting data, are also under way.  

I have several sets of membranes available for collaborative purposes: 
interested parties could either request a set of membranes, or 
alternatively, send me a small amount of DNA probe for hybridisation.  The 
positively-hybridising clones will be returned to the investigator, along 
with any other data known about the particular clone(s).

If the demand for sets of membranes is high, we may have to levy a *small* 
charge to cover the costs of materials etc.  We do not anticipate that the 
cost would be great.

In closing, I can heartily recommend this approach to genome mapping as it 
is relatively straight-forward, cost-effective, and usually quite rapid.  If 
there is anyone out there who might be interested in getting more 
information, do not hesitate to contact me by email at:

a.ivens at ic.ac.uk

and I will do my best to reply promptly.  I have requested subscription to 
this gopher service, so hopefully I'll be on-line shortly.

Looking forward to hearing from you!

Al Ivens

This messagette came from Dr. Al Ivens, Department of Biochemistry, Imperial 
College, Exhibition Road, London SW7 2AZ, U.K.

Currently the "lab" phone number is -XX-44-71-5895111 ext. 55220, my office 
is -XX-44-71-5945198, and the Fax number is -XX-44-71-2250960.

Hear from you soon!