摘要
以人血基因组DNA为模板 ,合成两段 2 0个寡聚核苷酸为引物 ,经过PCR扩增 ,得到 4 80bp的片段 ,克隆到pMD18-T载体中 ,经电泳、酶切、PCR鉴定后测定序列。序列分析表明所克隆的人端粒酶RNA(humantelomeaseRNA ,hTR)基因含有 4 80bp ,包括约 4 50bp的编码模板区序列和约 30bp的上游调控区序列 ,其中模板区的 11个核苷酸 ( 5’ -CUAACCCUAAC - 3’)合成端粒亚单位 (TTAGGG) n。结果与文献报道的人肾癌 2 93细胞系hTRcDNA序列完全相同 ,同源性达 10 0 % ,表明已经获得了hTR基因克隆。
The chromosome DNA of human blood as template,synthesis 20 oligonucleotides as primer,a fragment which contain 480 base pairs was amplified by PCR and cloned into vector pMD18-T and sequenced.The result of sequence analysis shows this hTR(human telomease RNA) gene fragment containing about 450bp complete template and about 30bp adjacent upstream region regulatory sequence was cloned.The template region of hTR encompasses 11 nucleotides (5'-CUAACCCUAAC-3') complementary to the human telomere sequence (TTAGGG) n.Compared with the known sequence of Feng's hTRcDNA from 293 cells,homology of nucleotide of hTR gene is 100%.So hTR gene has been got.
出处
《生物技术》
CAS
CSCD
2000年第6期5-8,共4页
Biotechnology
