Monochromosomal hybrids for the analysis of the human genome

Technical Report · Mon Jan 01 04:00:00 EST 1990

DOI:https://doi.org/10.2172/6038454· OSTI ID:6038454

Athwal, R S

In this research project the authors proposed to develop rodent/human hybrid cell lines each containing a single different human chromosome. The human chromosomes will be marked with Ecogpt and stably maintained by selection in the hybrid cells. The experimental approach to produce the proposed cell lines involve the following: they will first transfer a cloned selectable marker, Ecogpt (an E. coli gene for xanthine-guanine phosphoribosyltransferase: XGPRT) to normal diploid human cells using a retroviral vector. 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 they also use directed integration of Ecogpt into the chromosome present in rodent cells, otherwise not marked with a selectable marker. This allows them to complete the bank of proposed cell line. The human chromosome, since it will be marked with a selectable marker, can be transferred to any other cell line of interest for complementation analysis. Clones of each cell line, containing varying size 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 method will facilitate physical mapping and determination of gene order on a chromosome. 1 fig.

• Research Organization: University of Medicine and Dentistry of New Jersey, Newark, NJ (USA). Dept. of Microbiology and Molecular Genetics
• Sponsoring Organization: DOE/ER
• DOE Contract Number: FG02-89ER60866
• OSTI ID: 6038454
• Report Number(s): DOE/ER/60866-2; ON: DE91005006
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BACTERIA
BIOLOGICAL MARKERS
CELL CULTURES
CHROMOSOMES
CLONE CELLS
ENZYMES
ESCHERICHIA COLI
GENES
GENETIC ENGINEERING
GENETIC MAPPING
MAMMALS
MAN
MAPPING
MICE
MICROORGANISMS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PRIMATES
RODENTS
TRANSFERASES
VERTEBRATES