Nucleic Acids Research, 1993, Vol. 21, No. 8 1747-1752
© 1993
MOLECULAR BIOLOGY |
Cloning and characterization of a naturally occurring antisense RNA to human thymidylate synthase mRNA
Department of Experimental Therapeutics and The Grace Cancer Drug Center Roswell Park Cancer Institute, Buffalo, NY 14263, USA
Received February 9, 1993. Accepted March 11, 1993.
Based upon reverse transcription and polymerase chain reaction results with human KB cell RNA, a cDNA (i.e., 3'rTS1, 1557 nt) with complementarity to thymidylate synthase mRNA was cloned and sequenced. Northern blot analysis showed that 3'rTS1 corresponded to a cytoplasmlc 1.8 kb RNA found in several tumor cell lines. The remaining 5' region of this antisense RNA was cloned by a RACE (Rapid Amplification of cDNA Ends) procedure. A full length cDNA (I.e., rTS, 1811 nt) was generated by splicing 3'rTS1 with RACE-generated cDNA. rTS RNA is likely a mRNA that contains four open reading frames. Based upon sequence analysis of the RACE cDNAs and the rTS cDNA, rTS RNA is likely processed from a gene containing at least six introns. Northern blot analysis indicates rTS RNA is expressed in a variety of human tumor cell lines and an aberrant form Is expressed in a methotrexate-cell line.
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