Resource Development 



Abstracts 



Monochromosomal Hybrids for the Analysis 

 of the Human Genome 



Raghbir S. Athvval 



Department of Microbiology and Molecular Genetics. University of Medicine and 

 Dentistry of New Jersey. New Jersey Medical School. Newark. NJ 07103-2757 

 (201)456-4484 



In this research project we have proposed to develop rodent/human hybrid cell lines, 

 each containing a single different human chromosome. The human chromosomes will 

 be marked with Ecagpl and stably maintained by selection in the hybrid cells. 



This experimental approach to producing the proposed cell lines involves the following: 

 Using a retroviral vector, we will first transfer a cloned selectable marker. Ecogpt (an 

 E. coli gene for xanthine-guanine phosphoribosyltransferase: XGPRT), to normal 

 diploid human cells. The transferred gene will integrate at random into multiple sites in 

 the recipient cell genome. Clonal cell lines from independent transgenotes will each 

 carry the selectable marker integrated into a different site and perhaps a different 

 chromosome. The chromosome carrying the selectable marker will then be transferred 

 further to mouse cells by microcell fusion. In addition, we will use directed integration 

 of Ecogpt into the chromosome present in rodent cells, not otherwise marked with a 

 selectable marker. This will allow us to complete the bank of proposed cell lines. 



Since the human chromosome will be marked with a selectable marker, it can be 

 transferred to any other cell line of interest for complementation analysis/ Clones of 

 each cell line, containing varying sized segments of the same chromosome produced by 

 selection for the retention or loss of the selectable marker following X-irradiation or by 

 metaphase chromosome transfer, will facilitate physical mapping and determination of 

 gene order on a chromosome. 



52 



