一个新的C2H2型锌指蛋白cDNA基因的克隆

CLONING OF A NOVEL C2H2-TYPE ZINC FINGER cDNA GENE

目的 :从K5 6 2细胞中克隆新的锌指蛋白cDNA基因片段。方法 :提取K5 6 2细胞中总RNA ,逆转录后 ,用CX2C和H -CLink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K5 6 2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。

Objective:To clone a novel C2H2-type zinc finger cDNA gene,the total RNA isolated from the K562 cells was purified.Methods:The cNDA first strand was synthesized fro the total RNA,after reverse transcription,and the cDNA was amplified by add-on PCR.Products were digested with both EcoR I and BamH I and then were recombined into the pGEM7Zf(+) vector.Final,we measured five the inserted DNA fragments,and compared homology with nucleotide sequence database of GenBank.Results:One of them was a novel cDNA gene,which encoding product containing five C2H2-type zinc finger domain at least.Therfore.Conclusions:In our laboratory discovered the novel C2H2-type zinc finger domain cDNA gene laid a foundation for research of the total sequence and function of the gene.