Measurement of transcribed human X-chromosomal DNA sequences transferred to rodent cells by chromosome-mediated gene transfer

Journal Article · Fri Jan 01 04:00:00 EST 1982 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.2.1.52· OSTI ID:5606724

McBride, O W; Olsen, A S; Aulakh, G S; Athwal, R S

Transfer of genetic information can be effected by incubation of cultured eucaryotic cells with isolated metaphase chromosomes. In most cases, a resulting transformed cell contains only a fragment of a donor chromosome. The amount of transferred donor DNA has been quantified in 11 independent mouse A9 transformants by nucleic acid hybridization analysis. Each transformant had been selected for hprt (hypoxanthine phosphoribosyltransferase; EC 2.4.2.8) transfer and contained part of the human X chromosome. A labeled probe of transcribed human X-chromosomal DNA was prepared by hybridization of nick-translated unique-sequence human DNA with whole cellular RNA from a human-mouse hybrid line, A9/HRBC2-A, containing a single human chromosome, X. The amount of human X-chromosomal DNA in the transformants was quantitated by comparing the hybridization of this probe with transformant and A9/HRBC2-A DNAs. Two unstable transformants which had a microscopically detectable donor chromosome fragment contained 15% of the human C-chromosomal single-copy DNA. Four other unstable transformants contained 4 to 7% of human X-chromosomal DNA sequences. The transferred DNA was below the level of detection in three other unstable and in all three stable transformants. The authors conclude that the initial transfer event can introduce a substantial amount of genetic information but only smaller amounts of DNA are stably incorporated by integration.

• Research Organization: Dept. of Pharmacology, College of Medicine and Dentistry of New Jersey, Rutgers Medical School, Piscataway, NJ 08854
• OSTI ID: 5606724
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 2:1; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
CELL CULTURES
CELL DIVISION
CHEMICAL ANALYSIS
CHROMOSOMAL ABERRATIONS
CHROMOSOMES
CLONE CELLS
CODONS
DNA SEQUENCING
ENZYME ACTIVITY
ENZYMES
EVALUATION
GLYCOSYL TRANSFERASES
HETEROCHROMOSOMES
HYBRIDOMAS
HYPOXANTHINE PHOSPHORIBOSYLTRANSFERASE
LABELLING
MAMMALS
MAN
MITOSIS
MUTANTS
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PENTOSYL TRANSFERASES
PRIMATES
PROBES
QUANTITATIVE CHEMICAL ANALYSIS
RATS
RNA
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
TRANSCRIPTION
TRANSFER RNA
TRANSFERASES
VERTEBRATES
X CHROMOSOME