一个新的C2H2型锌指蛋白cDNA基因的克隆

摘要

To clone a novel C2H2-type zinc finger cDNA gene,the total RNA isolated from the K562 cells was purified.Methods:The cNDA firs t strand was synthesized fro the total RNA,after reverse transcription,and the c DNA was amplified by add-on PCR.Products were digested with both EcoR I and Bam H I and then were recombined into the pGEM7Zf(+) vector.Final,we measured five t he inserted DNA fragments,and compared homology with nucleotide sequence databas e of GenBank.Results:One of them was a novel cDNA gene,which enc oding product containing five C2H2-type zinc finger domain at least.Therfore.Conclusions:In our laboratory discovered the novel C2H2-type zi nc finger domai n cDNA gene laid a foundation for research of the total sequence and function of the gene.%从K562细胞中克隆新的锌指蛋白cDNA基因片段。方法：提取 K562细胞中总RNA，逆转录后，用CX2C和H-C Link引物进行加端PCR，扩增产物经限制性内切 酶 酶切后，重组至pGEMTZf(+)载体，用末端终止法测定插入片段的序列。结果：测出五个插入 片段的序列。与GenBank核酸数据库中的已知序列进行同源性比较分析，发现一个序列为新 的cDNA基因片段。该基因编码产物至少含有五个C2H2型锌指结构。结论：从K562细胞中克隆 出一个新的C2H2型锌指蛋白cDNA基因片段，为测定该基因的全序列和功能研究奠定了基础。