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- W3032850511 abstract "目的构建氯化血红素诱导性K562细胞抑制消减cDNA文库,以期鉴定出K562细胞中氯化血红素诱导性表达的珠蛋白合成调节因子cDNA基因.方法采用不同浓度的氯化血红素诱导培养K562细胞,经联苯胺阳性细胞百分率及血红蛋白浓度分析,选择其最佳诱导浓度.分别提取氯化血红素诱导培养的K562细胞(tester, 检测子)和未加诱导剂培养的K562细胞(driver, 驱赶子)mRNA,逆转录合成双链cDNA.经两轮消减杂交,两轮PCR扩增后,正向消减的PCR产物与T载体连接,构建K562细胞抑制消减cDNA文库,文库经蓝-白筛选后,纯化阳性克隆质粒,EcoRⅠ酶切及PCR扩增插入片段.结果以50μmol/L 氯化血红素诱导K562细胞,其联苯胺阳性细胞百分率及血红蛋白浓度均达到最大值,故选择50μmol/L 氯化血红素诱导培养K562细胞并构建K562细胞抑制消减cDNA文库.文库鉴定证实其阳性克隆中分别含有不同长度的插入片段.结论成功构建了氯化血红素诱导性K562细胞抑制消减cDNA文库,该消减cDNA文库结合生物信息学(Bioinformatics) 分析可用于深入研究K562细胞内氯化血红素诱导性表达基因的结构和功能." @default.
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- W3032850511 date "2003-01-20" @default.
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- W3032850511 title "Construction of Subtractive cDNA Library of K562 Cells" @default.
- W3032850511 doi "https://doi.org/10.3760/cma.j.issn.1008-1372.2003.01.002" @default.
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