genes can be used to detect repetitive, independent 
mutants containing the same sequence. 
DNA from C57BL/6 ovaries is being prepared, fol- 
low^ed by the amplification and cloning of all of the 
genes, both wild type and microrecombinant. 
Screening the I& clones for one or several particu- 
lar microrecombinants is accomplished -with a sin- 
gle oligonucleotide probe or a cocktail of oligonu- 
cleotide probes for known microrecombinant 
genes. 
The amplification of genes is accomplished 
using A*-specific oligonucleotide primers and the 
polymerase chain reaction (PGR). Using a series of 
PGR reactions. Dr. Geliebter's laboratory has been 
able to achieve a 10- to 100-million-fold amplifica- 
tion of the id' gene. The inclusion of restriction en- 
zyme sites on the oligonucleotide primers facilitates 
the cloning of the amplified product, to the extent 
that A* libraries with over 1 million clones are 
achievable. An elaborate scheme of gel purifications 
of the k!' gene, both before and after the PGR reac- 
tions, has been implemented, to avoid possible arti- 
facts inherent in the PGR. 
This amplification and cloning scheme should 
allow for the accurate determination of microre- 
combination frequencies of H-2 genes. These data 
will enable the comparison of microrecombinations 
in females versus males, different strains of mice, 
and different H-2 genes within the same mouse 
strain. Furthermore, the construction of donor 
gene libraries by the same procedures, followed by 
screening for if sequences, will determine if Qa 
and Tla region genes are recipients of H-2 DNA se- 
quences. This will help determine if the microre- 
combination process is reciprocal or nonreciprocal. 
Dr. Geliebter is also Assistant Professor and Uni- 
versity Fellow at The Rockefeller University. 
PUBLICATIONS 
Books and Chapters of Books 
Geliebter, J., Zeff, R.A., Spathis, R., Pfaffenbach, G., Nakagawa, M., McGue, B., Mashimo, H., Kesari, K., 
Hemmi, S., Hassenkrug, K., Borriello, R, Ajit Kumar, P., and Nathenson, S.G. 1988. The analysis of H-2 mu- 
tants: molecular genetics and structure/function relationships. In Major Histocompatibility Complex 
Genes and Their Role in Immune Function (David, G.S., Ed.). New York: Plenum, pp 169-176. 
Articles 
Geliebter, J., and Nathenson, S.G. 1988. Microrecombinations generate sequence diversity in the murine 
major histocompatibility complex: analysis of the KP"^^, j^m4^ j^hmio^ j^&wi7 mutants. Mol Cell Biol 
8:4342-4352. 
Hemmi, S., Geliebter, J. , Zeff, R.A., and Nathenson, S.G. 1988. Three spontaneous H-2^'' mutants are gener- 
ated by genetic micro-recombination (gene conversion) events. Impact on modulation of H-2-restricted 
immune responsiveness. J Exp Med 168:2319-2335. 
Neyt, G., Geliebter, J., Saloui, M., Morales, D., Meulemans, G., and Burny, A. 1989. Mutations located on both 
Fl and F2 subunits of the Newcastle disease virus fusion protein confer resistance to neutralization with 
monoclonal antibodies. J Virol 63:952-954. 
Sherman, D.R., Geliebter, J., and Cross, G.A.M. 1989. Rapid and simple amplification of a specific UNA se- 
quence by the polymerase chain reaction. Trends Genet 5:137. 
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