Worm Breeder's Gazette 8(2): 50

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Comparison of Tc1 Arrangement in High-Copy-Number Strains

D. Katzenberg, S. Emmons

We have analyzed the arrangement of Tc1 elements in several highcopy-
number strains.  Most of these strains originated from the C.  elegans 
laboratory in Lyon, France, and hence could be clonally related.  This 
group includes Bergerac (BO) (CGC), Bergerac (FR) (CGC), BL1 (formerly 
our 'Bergerac (SH)' from Bill Sharrock), and N62 (MRC).  One new wild 
isolate from California, DH424 (formerly EPC4), also has a large 
number of Tc1 elements in its genome.
We have found that Tc1 is arranged similarly in all these strains.  
We have used a two-dimensional system to visualize individual 
restriction fragments carrying Tc1 elements.  DNA was digested with 
one enzyme and fractionated on a sucrose gradient.  DNA from the upper 
one-third of the gradient was collected, digested with a second enzyme 
and analyzed for Tc1 elements on a Southern.  By this means about 80 
distinct bands were visualized.  The five strains analyzed had about 
70 fragments in common, and from five to 15 unique fragments.
Interestingly, DH424 was not distinct from the Bergerac group.  In 
fact, reckoning up similarities and differences, it was more closely 
related to FR, BL1 and N62 than to BO.  Either the amplification of 
Tc1 must have occurred before the separation of the DH424 and Bergerac 
lineages, or Tc1 has a strong preference for inserting at certain 
sites in the genome.
We also compared the structure of the Tc1 elements themselves in 
these strains and found no differences.  By doing a Southern after 
double digestion with Eco RV (which cuts within the inverted repeats 
20 base pairs from the ends of Tc1) and a second enzyme, it is 
possible to construct a consensus restriction map of genomic elements. 
In all these strains Tc1 elements are 1.6 kb in length and have 
similarly-placed SalI and XhoI sites.