人Peroxiredoxin6基因的克隆与鉴定

Cloning and identification of the human Peroxiredoxin 6 gene

目的构建pEGFP—N1-Prdx6真核表达载体，为进一步研究Peroxiredoxin6（Prdx6）在急性肺损伤中的作用奠定基础。方法利用RT—PCR方法扩增人Prdx6基因全长，测序证实后，将其定向亚克隆到真核表达载体pEGFP—N1中，提取质粒作双酶切和测序鉴定。结果人全长Prdx6基因序列正确插入pEGFP-N1载体，测序结果经BLAST分析与GenBank中报道的编码区cDNA序列完全一致。结论通过构建pEGFP-N1-Prdx6真核表达载体可以克隆人Prdx6基因。

Objective To construct and identify a vector carrying human Peroxiredoxin 6 （Prdx6） gene fused with enhanced green fluorescent protein （pEGFP-N 1） in order to study the function of Prdx6 gene in acute lung injury . Methods The full-length ofPrdx6 gene was obtained from A549 cell line by RT- PCR. After sequencing identification, the Prdx6 gene was inserted into eukaryotic expression vector pEGFP- N 1, then the plasmid was cut by restriction enzymes di- gestion and was identified by sequencing. Results The full-length coding sequence of Prdx6 was successfully inserted into the eukaryotic expression vector pEGFP-N 1, and the sequencing confirmation result showed that the sequence of Prdx6 cDNA was consistent with the cDNA sequences of the coding region in GenBank through BLAST analysis. Conclusion The human Prdx6 gene can be cloned via constructing the eukaryotic expression vector pEGFP- N 1- Prdx6.