The use of human repetitive DNA to target selectable markers into only the human chromosome of a human-hamster hybrid cell line (AL)
We used the plasmid BLUR-8 that contains an 800-base pair (bp) sequence of human repetitive Alu DNA in a cotransfection protocol to target the plasmids pSV2neo or EBO-pcD-leu-2 (hygro) into a single site of the sole human chromosome, number 11, of a Chinese hamster-human hybrid cell line (AL). The neo and hygro plasmids confer resistance to the antibiotics G418 and hygromycin, respectively. Of the 33 cotransfected clones with single-site insertions, 1/13 without BLUR-8 and 6/20 with BLUR-8 were only in human chromosome 11. A frequency of insertion of 1/13 is not different than expected by chance (rho = 0.3512). On the other hand, the probability that 6/20 insertions, as seen with BLUR-8, occurred by chance is low (rho = 0.0003). We suggest that the human DNA sequences contained in BLUR-8 targeted insertions into only the human chromosome.
Waldren
C
C
Radiological Health Sciences, Colorado State University, Fort Collins 80523.
Braaton
M
M
Vannais
D
D
Fouladi
B
B
Parker
R D
RD
D000818:Animals; D015139:Blotting, Southern; D002880:Chromosomes, Human, Pair 11; D006224:Cricetinae; D004560:Electricity; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D006822:Hybrid Cells; D010957:Plasmids; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection