Dolnick B J
Department of Experimental Therapeutics, Roswell Park Cancer Institute, Buffalo, NY 14263.
Nucleic Acids Res. 1993 Apr 25;21(8):1747-52. doi: 10.1093/nar/21.8.1747.
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 cytoplasmic 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 from is expressed in a methotrexate-cell line.
基于对人KB细胞RNA进行逆转录和聚合酶链反应的结果，克隆并测序了与胸苷酸合成酶mRNA互补的cDNA（即3'rTS1，1557个核苷酸）。Northern印迹分析表明，3'rTS1对应于在几种肿瘤细胞系中发现的一种1.8 kb细胞质RNA。通过RACE（cDNA末端快速扩增）方法克隆了这种反义RNA的其余5'区域。通过将3'rTS1与RACE产生的cDNA拼接，产生了全长cDNA（即rTS，1811个核苷酸）。rTS RNA可能是一种包含四个开放阅读框的mRNA。基于RACE cDNA和rTS cDNA的序列分析，rTS RNA可能是由一个至少包含六个内含子的基因加工而来。Northern印迹分析表明，rTS RNA在多种人肿瘤细胞系中表达，并且在甲氨蝶呤抗性细胞系中表达一种异常形式。
