小鼠ZP2肽的基因克隆和原核表达载体的构建

Gene cloning of mouse ZP2 peptide and construction of its expression vector

目的:克隆小鼠卵透明带2(mZP2)肽的基因并构建原核表达载体。方法:从小鼠卵巢组织中分离出mRNA,并以此作为模板,通过RT-PCR扩增出小鼠ZP2肽的cDNA片段,将其克隆到pET原核表达载体。将该重组mZP2基因转化Rosetta-gami(DE3)pLysS菌,通过SDS-PAGE和Western blotting分析重组蛋白的表达情况。结果:克隆小鼠ZP2肽的cDNA片段,构建原核表达载体pET-mZP2,经SDS-PAGE和Western blotting鉴定表明表达产物是约36 ku具有6个组氨酸标签肽的融合蛋白。结论:成功克隆小鼠ZP2基因片段,构建的mZP2原核表达载体具有表达功能。

Aim：To clone the gene of mouse ZP2 peptide and to construct a prokaryotic expression vector of mZP2.Methods： The total RNA was extracted from mouse ovarian tissue.The ZP2 cDNA fragment was amplified by RT-PCR,and inserted into the MCS of vector pET for induced expression in Rosetta Bacterium.The recombinant mZP2-His tag fusion protein was separated by SDS-PAGE and identified by Western blotting.Results： mZP2 cDNA fragment was cloned and linked to expression vector pET.The recombinant mZP2 fusion protein was detected in the lysate of pET-mZP2-transformed Rosetta bacteria by SDS-PAGE and Western blotting.Conclusion：The mZP2 cDNA fragment has been cloned and recombinant expression vector pET-mZP2 has been constructed with functions.