CLONING AND CHARACTERIZATION OF A NATURALLY-OCCURRING ANTISENSE RNA TO HUMAN THYMIDYLATE SYNTHASE MESSENGER-RNA
CLONING AND CHARACTERIZATION OF A NATURALLY-OCCURRING ANTISENSE RNA TO HUMAN THYMIDYLATE SYNTHASE MESSENGER-RNA
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DOI：
10.1093/nar/21.8.1747
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发表时间：
1993-04-25
影响因子：
14.9
通讯作者：
DOLNICK, BJ
中科院分区：
文献类型：
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作者：
DOLNICK, BJ
根据人 KB 细胞 RNA 的逆转录和聚合酶链反应结果，克隆并测序了与胸苷酸合酶 mRNA 互补的 cDNA（即 3'rTS1，1557 nt）。 Northern 印迹分析表明，3'rTS1 对应于在几种肿瘤细胞系中发现的细胞质 1.8 kb RNA。通过 RACE（cDNA 末端快速扩增）程序克隆该反义 RNA 的剩余 5' 区域。通过将 3'rTS1 与 RACE 生成的 cDNA 拼接来生成全长 cDNA（即 rTS，1811 nt）。 rTS RNA 可能是包含四个开放阅读框的 mRNA。根据 RACE cDNA 和 rTS cDNA 的序列分析，rTS RNA 可能是由包含至少六个内含子的基因加工而成。 Northern印迹分析表明rTS RNA在多种人类肿瘤细胞系中表达，并且异常形式在甲氨蝶呤细胞系中表达。
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 form is expressed in a methotrexate-cell line.