中文摘要：

利用表达载体pIRES-NA研究H5N1型神经氨酸酶（neuramidinase,NA）基因在NIH-3T3细胞中表达,为今后转基因家禽的培育提供证据。通过构建重组表达载体转染细胞经G418筛选后,利用PCR方法检测NA基因在细胞基因组中整合,利用RT-PCR和Western Blotting的方法分别在RNA及蛋白质水平上检测NA基因在细胞中表达。结果表明细胞转染经筛选后,提取细胞基因组进行PCR,能扩增出NA基因片段,约1.4kb;同时RT-PCR能检测到NA基因mRNA,Western Blotting能检测到细胞表达约55ku NA蛋白,构建重组载体能整合到NIH-3T3细胞基因组中,NA基因能在细胞中表达。

英文摘要：

The aim of this experiment was to study the expression of H5N1 subtype neuramidinase gene transferred to NIH-3T3 cells by constructing recombinant expression vector pIRES-NA in order to provide the proof of producting transfer-gene fowls further.After cells were transfected with pIRES-NA and sieved with G418,the PCR method was used to examine whether the pIRES-NA vector was integrated to the cell gene set or not;the methods of the RT-PCR and the Western Blotting were respectively used to examine the expression of NA gene in cells transcripted and translated to the protein.The result showed that after cell transfection and sieving,cell gene set was withdrawn and amplified NA gene （1.4 kb） by PCR;the RT-PCR could examine mRNA of NA gene in the cells ;NA protein ,about 55 ku,from the cells could be examined by Western blotting.As a conclusion,recombinant expression vector pIRES-NA could be integrated to the cell gene set,and NA gene might be transcripted and translated to protein in the cells.